Anti-GFAP 抗体 [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric)

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

IHC image of GFAP staining in a section of frozen Human normal cerebral cortex performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab323239, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Composite multiplex immunofluorescence staining of Iba1, GFAP and MAP2 staining in a section of formalin-fixed paraffin-embedded human cerebral cortex*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 5µg/ml dilution (shown in green), ab300645 at 5µg/ml (shown in magenta), and ab178846 at 5µg/ml (shown in yellow). Then incubated for 1 hour with ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed 1/1000, ab150084 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed 1/1000, and ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

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

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Immunofluorescence staining of GFAP in sections of formalin-fixed paraffin-embedded human cerebral cortex*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9.0) for 40 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Negative control : confocal image showing no staining on a section of frozen normal Human liver performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab323239, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Negative Control : PC-12. Flow cytometry (intracellular) analysis of PC-12 (rat adrenal gland pheochromocytoma cell) cells fixed with 4% PFA showing no staining of GFAP with ab323239 used at 1/5000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

ab323239 showing no staining of GFAP in NIH/3T3 (mouse embryonic fibroblast) cells at 1/50 dilution (10 ug/mL). The cells were fixed with 4% Paraformaldehyde, and permeabilized with 0.1% PBS-Triton X-100. Negative control : confocal image showing no staining in NIH/3T3. The counterstain, ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1/1000 (1 ug/mL) dilution was observed in magenta. ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000 (2 μg/ml) was used as secondary antibody. Nuclear DNA was labelled with DAPI (shown in blue).

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

-ve control 1 : ab323239 (Chicken antibody), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594).

-ve control 2 : GFAP (Mouse antibody), followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed.

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Negative control : confocal image showing no staining on a section of Rat liver (perfusion fixed, 4% solution PFA).

The section was incubated with ab323239 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Immunohistochemical analysis of Mouse hippocampus (perfusion fixed, 4% solution PFA) labeling GFAP with ab323239.

Panel A : merged staining of anti-GFAP using ab323239 (green), counterstained with anti-NeuN ab190565 (magenta) on mouse hippocampus.

Panel B : anti-GFAP stained on mouse hippocampus.

Panel C : anti-NeuN stained in neurons of mouse hippocampus.

The section was incubated with ab323239 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Immunohistochemical analysis of Rat hippocampus (perfusion fixed, 4% solution PFA) labeling GFAP with ab323239.

Panel A : merged staining of anti-GFAP ab323239 (green), counterstained with anti-NeuN ab190565 (magenta) on rat hippocampus.

Panel B : anti-GFAP stained on rat hippocampus.

Panel C : anti-NeuN stained in neurons of rat hippocampus.

The section was incubated with ab323239 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). The section was then mounted using Fluoromount®.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

ab323239 staining GFAP in mouse primary neural/glia cells at 1/50 dilution (10 ug/mL). The cells were fixed with 4% Paraformaldehyde, and permeabilized with 0.1% PBS-Triton X-100. Confocal image showing positive cytoplasmic staining in mouse astrocytes (shown in green). The counterstain, Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody at 1/100 dilution was observed in magenta. ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000 (2 μg/ml) was used as secondary antibody. Nuclear DNA was labelled with DAPI (shown in blue).

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

-ve control 1 : ab323239 (Chicken antibody), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594).

-ve control 2 : GFAP (Mouse antibody), followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed.

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Negative control : confocal image showing no staining on a section of Mouse liver (perfusion fixed, 4% solution PFA).

The section was incubated with ab323239 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

ab323239 staining GFAP in rat primary neural/glia cells at 1/50 dilution (10 ug/mL). The cells were fixed with 4% Paraformaldehyde, and permeabilized with 0.1% PBS-Triton X-100. Confocal image showing positive cytoplasmic staining in rat astrocytes (shown in green). The counterstain, Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody at 1/100 dilution was observed in magenta. ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000 (2 μg/ml) was used as secondary antibody. Nuclear DNA was labelled with DAPI (shown in blue).

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

-ve control 1 : ab323239 (Chicken antibody), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594).

-ve control 2 : GFAP (Mouse antibody), followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

ab323239 showing no staining of GFAP in PC-12 (rat adrenal gland pheochromocytoma cell) cells at 1/50 dilution (10 ug/mL). The cells were fixed with 4% Paraformaldehyde, and permeabilized with 0.1% PBS-Triton X-100. Negative control : confocal image showing no staining in PC-12. The counterstain, ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1/1000 (1 ug/mL) dilution was observed in magenta. ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000 (2 μg/ml) was used as secondary antibody. Nuclear DNA was labelled with DAPI (shown in blue).

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

-ve control 1 : ab323239 (Chicken antibody), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594).

-ve control 2 : GFAP (Mouse antibody), followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Immunofluorescence staining of GFAP in sections of formalin-fixed paraffin-embedded rat brain (positive) and rat skeletal muscle (negative).

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

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

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Immunofluorescence staining of GFAP in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse skeletal muscle (negative).

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

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

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Negative Control : NIH/3T3. Flow cytometry (intracellular) analysis of NIH/3T3 (mouse embryonic fibroblasts) fixed with 4% PFA showing no staining of GFAP with ab323239 used at 1/50000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Flow cytometry (intracellular) analysis of Mouse primary neural/glia cells fixed with 4% PFA staining GFAP with ab323239 used at 1/50000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Flow cytometry (intracellular) analysis of Rat primary neural/glia cells fixed with 4% PFA staining GFAP with ab323239 used at 1/5000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.

• WB

Lab

Western blot - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (AB323239)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602).

Negative control : liver.

All lanes:

Western blot - Anti-GFAP antibody [EPR1034Y] - Astrocyte marker - Chicken IgY (Chimeric) (ab323239) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/10000 dilution

Observed band size: 45-50 kDa

false

Exposure time: 3min