AB199093

Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control

RabMAb
Recombinant
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(30 Publications)

Anti-Rabbit IgG, monoclonal [EPR25A] - Isotype Control- Alexa Fluor® 647 conjugated (ab199093) is a rabbit recombinant monoclonal antibody used in Flow Cytometry, ICC/IF.

- Biophysical QC for unrivalled batch-batch consistency
- Over 10 publications

41 Images

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199093 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-87 MG(human glioblastoma-astrocytoma epithelial cell) cells labelling Ataxin 3 with ab315423 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labelling Integrin alpha 4/CD49D with ab316939 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

human PBMC are co-stained with CD3 conjugated Alexa Fluor®488.
Gated on viable cell.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CLEC12A with ab317063 at 1/5000 dilution (0.01ug)/ Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Gated on viable cells. Cells were co-stained with anti-CD3 conjugated to Alexa Fluor® 488.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labelling ILT-4 with ab313898 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Cells are co-stained with CD14 conjugated to BV510. Gated on viable cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD45 with ab315960 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Negative control : 293T. Gated on viable cells.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 μg/ml PHA for 18h (bottom), either stained with ab196847 (right) or isotype control (left). The cells were fixed using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab196847) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Huh7 (human hepatocellular carcinoma epithelial cell) treated with 300nM thapsigargin for18hours (Red) / Untreated Huh7 (Dotted Red) cells labelling XBP1 with ab317358 at 1/5000 dilution(0.01ug)/Red and Dotted Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with myc-tagged IL-23R expression vector containing a Myc tag (Middle) / 293T cells transfected with an empty vector containing a Myc tag (Right) cells labelling IL-23R with ab319084 at 1/5000 dilution (0.01ug) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody) (Blue).

Live cells were surface stained with ab319084, then fixed with 2% PFA followed by intracellular staining with Myc-tag conjugated to Alexa Fluor® 488.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 µg/ml PHA for 18h (bottom), either stained with ab326136 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 µg/ml human IgG, 10% normal goat serum and BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (1/10) to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab326136) (1x 106 in 100µl at 0.2 µg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). The cells were simultaneously stained with CD3. Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type Jurkat (green line) and IDH2 knockout Jurkat stained with ab326447 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326447) (1x 106 in 100μl at 0.0016 μg/ml (1/312500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in Jurkat WT cells (black line) and Jurkat-IDH2 KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 μg/ml PHA for 18h (bottom), either stained with ab195020 (right) or isotype control (left). The cells were fixed using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab195020) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa(human cervical adenocarcinoma epithelial cell) cells labelling JNK1 + JNK2 + JNK3 with ab315842 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of K-562 (human chronic myelogenous leukemia lymphoblast, Left)/Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling Integrin alpha 4/CD49D with ab316939 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cell.
Negative control : K-562 (PMID : 16105875).

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab197864 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab197864) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing left wild-type HAP1 positive cells and right negative FASN knockout HAP1 stained with ab319690 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab319690) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500 dilution)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast) cells labelling IL-10 with ab305044 at 1/500 dilution (0.1ug) (Red) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab198587 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab198587, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor^® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / 293T cells transfected with a mouse CCR4 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling CCR4 with ab324334 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Cells were surface stained with isotype control or our antibody. Then fixed with 2% PFA for 10min followed by intracellularly stained with anti-myc tag conjugated to Alexa Fluor® 488.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a mouse CD27 expression vector (Middle) / 293T cells transfected with an empty expression vector (Right) cells labelling CD27 with ab324178 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were co-stained with Myc tag conjugated to Alexa Fluor®488.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Isotype (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with Myc-tagged ENPP3 overexpression construct (Middle) / 293T transfected with an empty vector containing a myc-His tag (Right) cells labelling ENPP3/B10 with ab319126 at 1/500 dilution (0.1ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody) (Blue).

Cells were co-stained with anti-myc conjugated to Alexa Fluor 488.
Gated on viable cells.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype / 293T (human embryonic kidney epithelial cell) transfected with a TRDC (human TCR delta) protein expression vector containing a myc-His tag(Up Right) / 293T transfected with a TRGC1 protein (human TCR gamma) expression vector(Down Left) / 293T transfected with an empty vector containing a myc-His tag(Down Right) cells labelling TCR delta with ab317716 at 1/500 dilution (0.1ug)/ Up Right, Down Left and Down Right (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HeLa (green line) and GYS1 knockout HeLa stained with ab326127 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326127) (1x 106 in 100µl at 0.008 µg/ml (1/62500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HeLa WT cells (black line) and HeLa-GYS1 KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.This antibody gave a positive signal in wild-type HeLa fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / Daudi (human Burkitt's lymphoma lymphoblast, Middle) / K-562 (human chronic myelogenous leukemia lymphoblast, Right) cells labelling MICB with ab325223 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cells.
Negative control : 293T, Daudi (PMID : 28154561).

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing left wild-type HAP1 positive cells and right negative ACLY knockout HAP1 stained with ab205430 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab205430) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HEK293T (green line) and HK1 knockout HEK293T stained with ab197864 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab197864) (1x 10⁶ in 100μl at 5 μg/ml (1/100)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HEK293T WT cells (black line) and HEK293T-HK1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab326179 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab326179) (1x 106 in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3. Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte, Left) and Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD3D with ab198937 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor^® 647) (ab199093) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Negative control : THP-1.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 1%FBS for 3hours then 100nM Calyculin A for 10min (Red) / Untreated control (Dotted Red) cells labelling IKB alpha (phospho S36) with ab317485 at 1/5000 dilution (0.01ug) /Red and Dotted Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) treated with 500ng/ml ionomycin and 10ng/ml PMA for 3 hours (Lower left and right) / Untreated control (Upper left and right) cells labelling CD69 with ab325108 at 1/500 dilution (0.1ug) / Upper right and Lower right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control.

Cells were stained with anti-CD4 conjugated to Brilliant Violet 421.
Gated on viable lymphoid Cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized hela(human cervical adenocarcinoma epithelial cell) cells labelling Ataxin 3 with ab315423 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and PRDX2 knockout HAP1 stained with ab197041 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab197041) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500 dilution)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-PRDX2 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab214257 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab214257) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells transfected with a Red fluorescent protein expression vector containing a His-tag (upper) / 293T cells transfected with an empty vector containing a His tag (low) cells labelling RFP with ab323758 at 1/500 dilution (0.1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Cells are co-stained with Myc tag conjugated to Alexa Fluor®488.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Hela (human cervical adenocarcinoma epithelial cell, Left)/U-937 (human histiocytic lymphoma monocyte, Right) cells labelling CD18 with ab316944 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cell.
Negative control : HeLa

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) / HEL (human erythroleukemia erythroblast, Right) cells labelling VEGF Receptor 3 with ab313899 at 1/50 dilution (1 ug)/Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : MCF7. Gated on viable cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199093)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD18 with ab316944 at 1/500 dilution (0.1ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647) (ab199093) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD4 conjugated BV421.
Gated on viable cell.

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EPR25A

アイソタイプ

IgG

標識

Alexa Fluor® 647

励起波長/蛍光波長

Ex: 650nm, Em: 665nm

キャリアフリー

交差種

Human

アプリケーション

ICC/IF, IHC-P, Flow Cyt, Flow Cyt (Intra)

applications

特異性

Please note Abcam have optimised the validation of this product. In our hands, we observe an increase in background signal intensity with the use of Triton X-100 and would recommend using an alternative permeabilisation method such as methanol or saponin.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCyt-species-checked": "guaranteed", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.", "FlowCytIntra-species-checked": "", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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製品の詳細

Product Specifications
Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199093) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt and ICC/IF .

Quality and Validation
Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199093) has been cited over 19 times in peer reviewed journals and is trusted by the scientific community.
Flourescent conjugated antibodies are ideal tools for multiplex IHC and flow cytometry experiments.
Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199093) has 19 independent reviews from customers.
Alexa Fluor^® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199093) specifically detects rabbit IgG and is sold in 100 µL selling sizes.

Related Products
Conjugation-ready, carrier free format available for antibody clone (EPR25A) - ab210849.

Immunogen
Chemical / Small Molecule conjugated to keyhole limpet haemocyanin. KLH is a copper containing oxygen carrier occurring freely dissolved in the hemolymph of many molluscs and arthropods. KLH forms a large complex composed of ~50 kDa subunits.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA

出荷温度

Blue Ice

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Avoid freeze / thaw cycle|Store in the dark

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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

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文献 (30)

Recent publications for all applications. Explore the full list and refine your search

eLife 13: PubMed40996791

2025

Steady-state neuron-predominant LINE-1 encoded ORF1p protein and LINE-1 RNA increase with aging in the mouse and human brain.

Applications

Unspecified application

Species

Unspecified reactive species

Tom Bonnifet,Sandra Sinnassamy,Olivia Massiani-Beaudoin,Philippe Mailly,Heloise Monnet,Damarys Loew,Berangere Lombard,Nicolas Servant,Rajiv L Joshi,Julia Fuchs

PLoS pathogens 21:e1013087 PubMed40627772

2025

In vitro HIV DNA integration in STAT3 drives T cell persistence-A model of HIV-associated T cell lymphoma.

Applications

Unspecified application

Species

Unspecified reactive species

Michael Rist,Machika Kaku,John M Coffin

Journal of virology 99:e0220224 PubMed40152594

2025

Role of N-linked glycosylation sites in human ACE2 in SARS-CoV-2 and hCoV-NL63 infection.

Applications

Unspecified application

Species

Unspecified reactive species

Sabrina Noettger,Fabian Zech,Rayhane Nchioua,Chiara Pastorio,Christoph Jung,Timo Jacob,Steffen Stenger,Frank Kirchhoff

Nature communications 16:813 PubMed39827271

2025

α-Synuclein fibrils enhance HIV-1 infection of human T cells, macrophages and microglia.

Applications

Unspecified application

Species

Unspecified reactive species

Lia-Raluca Olari,Sichen Liu,Franziska Arnold,Julia Kühlwein,Marta Gil Miró,Ajeet Rijal Updahaya,Christina Stürzel,Dietmar Rudolf Thal,Paul Walther,Konstantin M J Sparrer,Karin M Danzer,Jan Münch,Frank Kirchhoff

Nucleic acids research 52:13561-13576 PubMed39569586

2024

Single-stranded DNA with internal base modifications mediates highly efficient knock-in in primary cells using CRISPR-Cas9.

Applications

Unspecified application

Species

Unspecified reactive species

Karen L Kanke,Rachael E Rayner,Jack Bozik,Eli Abel,Aparna Venugopalan,Ma Suu,Reza Nouri,Jacob T Stack,Gongbo Guo,Tatyana A Vetter,Estelle Cormet-Boyaka,Mark E Hester,Sriram Vaidyanathan

Molecular therapy. Nucleic acids 35:102339 PubMed39398224

2024

DNA-PKcs inhibition improves sequential gene insertion of the full-length cDNA in airway stem cells.

Applications

Unspecified application

Species

Unspecified reactive species

Jacob T Stack,Rachael E Rayner,Reza Nouri,Carlos J Suarez,Sun Hee Kim,Karen L Kanke,Tatyana A Vetter,Estelle Cormet-Boyaka,Sriram Vaidyanathan

JCI insight 9: PubMed39078714

2024

HIV-1 latency reversal and immune enhancing activity of IL-15 is not influenced by sex hormones.

Applications

Unspecified application

Species

Unspecified reactive species

Carissa S Holmberg,Callie Levinger,Marie Abongwa,Cristina Ceriani,Nancie M Archin,Marc Siegel,Mimi Ghosh,Alberto Bosque

Cell reports methods 3:100465 PubMed37323577

2023

Engineering CpG island DNA methylation in pluripotent cells through synthetic CpG-free ssDNA insertion.

Applications

Unspecified application

Species

Unspecified reactive species

Joshua Tompkins,Elizabeth Lizhar,Alireza Shokrani,Xiwei Wu,Jordan Berley,Diba Kamali,Deborah Hussey,Jonas Cerneckis,Tae Hyuk Kang,Jinhui Wang,Walter Tsark,Defu Zeng,Swetha Godatha,Rama Natarajan,Arthur Riggs

Advanced science (Weinheim, Baden-Wurttemberg, Germany) 10:e2300244 PubMed36938863

2023

Early Endosomes Act as Local Exocytosis Hubs to Repair Endothelial Membrane Damage.

Applications

Unspecified application

Species

Unspecified reactive species

Nikita Raj,Lilo Greune,Martin Kahms,Karina Mildner,Rico Franzkoch,Olympia Ekaterini Psathaki,Thomas Zobel,Dagmar Zeuschner,Jürgen Klingauf,Volker Gerke

Respiratory research 24:24 PubMed36691012

2023

RAGE inhibition alleviates lipopolysaccharides-induced lung injury via directly suppressing autophagic apoptosis of type II alveolar epithelial cells.

Applications

Unspecified application

Species

Unspecified reactive species

Xi Xiong,Jiaying Dou,Jingyi Shi,Yuqian Ren,Chunxia Wang,Yucai Zhang,Yun Cui

View all publications

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control