Product name

Anti-GFAP antibody [GF5]

Description

Mouse monoclonal [GF5] to GFAP

Host species

Mouse

Specificity

There is no cross-reactivity with other neurospecific proteins.

Tested applications

Suitable for: IHC-P,WB,ELISA,IHC-FoFr,IHC-Fr,ICC/IF,Flow Cyt,ICC

Species reactivity

Reacts with: Mouse, Rat, Human

Immunogen

This clone has been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunised with purified glial fibrillary acidic protein from human brain.

Positive control

• normal adult rat brain: lateral ventricle IHC-P: FFPE human hippocampus normal. IHC-P: FFPE rat brain normal.

General notes

Concentration varies from lot to lot and can be provided on request.

IHC image of GFAP staining in a formalin fixed, paraffin embedded rat normal brain tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10062 at 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.

 

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.

Mouse monoclonal to GFAP [GF5] (ab10062) was used in fixed murine cultures (mixed neurons/glia) at 1/100 overnight at 4°C. A secondary goat anti-mouse antibody was used for detection (Alexa Fluor 568; 1/400). Microscopy revealed diffuse cytosolic labelling. Coounterstaining with TO-PRO-3 (Molecular Probes; 660nm (converted here to blue colour) was used to identify the nucleus. The “fibrous” anti-GFAP staining of murine mixed cultures is typical of what is expected.

Overlay histogram showing SH-SY5Y cells stained with ab10062 (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 (ab10062, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

IHC image of GFAP staining in a formalin fixed, paraffin embedded human normal hippocampus tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab10062 at 1/500 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.

 

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.

 

GFAP antibody [GF5] - Astrocyte Marker (ab10062) immunocytochemical detection in stimulated Cor1 cells. Stimulated Cor1 cells were fixed in formaldehyde, permeabilized, blocked in 1% BSA for 10 mins @ rt°C. Primary Antibody ab10062 incubated at 1/1500 for 2 hours in TBS/BSA/azide/0.3% triton. Secondary Antibody: anti mouse IgG Conjugated to: Alexa Fluor® 488 (1/1000).

See Abreview

GFAP antibody [GF5] - Astrocyte Marker (ab10062; 1/250 for 16h) used in Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) on Rat Tissue sections (adult brain: lateral ventricle showing astrocytes).Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Secondary Antibody: biotin labelled goat anti mouse Ig (1/200). This image shows the exit point for the progenitor olfactory neurones, of the lateral ventrical subventricular zone.

Mouse monoclonal to GFAP [GF5] (ab10062) was used in fixed rat glial cultures at a dilution of 1/100, and incubated overnight at 4°C. Alexa Fluor 568 (1/400) secondary goat anti-mouse antibody was used for detection. Fluoresence microscopy revealed diffuse cytosolic labelling. Counterstaining with TO-PRO-3 (Molecular Probes; 660nm (converted here to blue colour) was used to identify the nucleus. The “fibrous” anti-GFAP staining of murine mixed cultures is typical of what is expected.