FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with HEK293T-Myc-His-hSNAI1 vector labeling 6X His tag® with ab318311 (middle panel) at 1.0μg/ml (1/500) compared with a Rabbit IgG [EPR25A] FITC (left panel) Gate is set between transfected and untransfected HEK-293T cells.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab314949 (right) or Rabbit IgG [EPR25A] FITC (left). PBMCs were incubated for 30 min on ice 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 ab314949 or Rabbit IgG [EPR25A] FITC (1x 10⁶in 100 μl at 0.2 μg/ml (1/2500)) for 30min on ice. The cells were simultaneously stained with CD19.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable lymphocytes.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of 293T cells transfected with a Red fluorescent protein expression vector containing a Myc-His-tag (upper right) / 293T cells transfected with an empty vector containing a Myc-His-tag (lower right) with ab323681 or 293T cells transfected with a Red fluorescent protein expression vector containing a Myc-His-tag (upper left) / 293T cells transfected with an empty vector containing a Myc-His-tag (lower left) with Rabbit IgG monoclonal [EPR25A] - Isotype Control (FITC) (ab223339). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 90% methanol for 30 min. Cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab323681) (1x 10⁶ in 100μl at 1 μg/ml (1/500)) for 30min at 22°C.

Isotype control antibody Rabbit IgG monoclonal [EPR25A] - Isotype Control (FITC) (ab223339) was used at the same concentration and conditions as the primary antibody. The cells were simultaneously stained with Myc.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab314951 (right) or Rabbit IgG [EPR25A] FITC (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab314951 or Rabbit IgG [EPR25A] FITC (1x 10⁶in 100 μl at 1.0 μg/ml (1/500)) for 30min on ice. The cells were simultaneously stained with CD19.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable lymphocytes.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry overlay histogram showing RAW264.7 treated with 100ng/ml LPS for 3h, then together with 300ng/ml BFA for another 3h (green line) or untreated RAW264.7 stained with ab323680 (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 incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab323680) (1x 10⁶ in 100μl at 1 μg/ml (1/500)) for 30min at 22°C.

Isotype control antibody Rabbit IgG [EPR25A] FITC was used at the same concentration and conditions as the primary antibody (RAW264.7 treated - black line, RAW264.7 untreated - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab315141 (right) or Rabbit IgG [EPR25A] FITC (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab315141 or Rabbit IgG [EPR25A] FITC (1x 10⁶in 100 μl at 0.2 μg/ml (1/2500)) for 30min on ice. The cells were simultaneously stained with CD49b.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry overlay histogram showing left BEND.3 positive cells and right negative NIH/3T3 stained with ab314950 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab314950) (1x 10⁶in 100μl at 1.0 μg/ml (1/500)) for 30min on ice.

Isotype control antibody (black line) was Rabbit IgG [EPR25A] FITC 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 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab323684 (right) or Rabbit IgG [EPR25A] FITC (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab323684 or Rabbit IgG [EPR25A] FITC (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab315141 (right) or Rabbit IgG [EPR25A] FITC (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab315141 or Rabbit IgG [EPR25A] FITC (1x 10⁶in 100 μl at 0.2 μg/ml (1/2500)) for 30min on ice. The cells were simultaneously stained with CD19.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Immunofluorescent analysis of HeLa (human cervical cancer) 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 ab223339 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control (FITC)) at 1/100 dilution (showing no signal) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (pseudocolored in red). Nuclear DNA was labelled 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 Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB223339)

Overlay histogram showing HeLa cells stained with ab139565 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab139565, 1/50 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) [EPR25A] FITC (ab223339) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.