GFAP antibody [GF5] - Astrocyte Marker (ab10062)

All antibodies work in different combinations of applications, species and samples, so we encourage our customers to write a short Abreview, so that future customers can make an informed decision about the usefulness of a particular antibody for their specific experiments.

Submit an Abreview and earn Abpoints

review by Carl Hobbs submitted on 15 April 2009
Application:	Immunocytochemistry/ Immunofluorescence
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Mouse Cell (Stimulated Cor1 cells)
Specification	Stimulated Cor1 cells
Fixative:	Formaldehyde
Permeabilization:	Yes - TBS/BSA/azide/0.3% Triton X
Blocking step:	BSA as blocking agent for  10 mins  · Concentration: 1 % · Temperature: rt°C
Primary Antibody
Dilution	1/1500
Incubation time:	2 hours  Diluent: TBS/BSA/azide/0.3% Triton
Secondary Antibody
Name:	Non-Abcam Antibody was used: anti mouse IgG Conjugation: Alexa Fluor® 488
Dilution:	1/1000
Additional Data
Rating	Excellent
Additional notes	I co- incubated stimulated Cor1 cells with Abcam's and a competitor anti-GFAP Abs: after washing in TBS x3 for 15minutes I then incubated the cells with buffer containing Goat anti Rabbit -Alexa 594, Goat anti Mouse Alexa- 488 and Hoechst for 1 hr at RT diluted in same buffer solution as the primary Abs. After further TBS washes, coverslips were picked up,rinses in distilled water performed to get rid of buffer salts and then the coverslips were mounted on glass slides, using a homemade "anti-fade" hard-setting mountant. Images were captured using a Zeiss Apotome Microscope system: Z-stacks of each channel were flattened in "cut-view". Adobe's Photoshop was used to merge red/greenchannels with Hoechst and then to globally enhance the composite image, using "unsharp mask". Seems to me that both Abs are co-localised. I did this because, in FFPW sections, the positivity seen with these Abs in the svz of mouse/rat brain FFPW sections of lateral ventricle is not identical (in my hands). My point therefore, is that one must always test several Abs raised against any particular protein unless Peer review concludes that one particular Ab reagent is agreed to be specific for any cell/tissue preparation. The cells were seeded/grown on coverslips and then fixed by Philipp Suetterlin, PhD student Wolfson Centre for Age-Related Diseases, Kings College London ...thanks, Phil! All previous Abreviews using Cor1 cells were possible only because of Phil's excellent skills in growing/maintaining/fixing Cor1 cells.
Image	GFAP antibody [GF5] - Astrocyte Marker for ICC/IF (Mouse) Copyright image used under license

Anonymous Abreview submitted on 29 April 2008
Application:	Immunocytochemistry
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Rat Cultured Cells (BMSC)
Specification	BMSC
Fixative:	Paraformaldehyde
Blocking step:	Serum as blocking agent for  30 mins  · Concentration: 10 % · Temperature: 20°C
Primary Antibody
Dilution	1/50
Incubation time:	48 hours  Temperature: 4°C Diluent: PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti mouse IgG rhodamine Conjugated Conjugation: Rhodamine
Dilution:	1/20
Additional Data
Rating	Average

review by Carl Hobbs submitted on 4 December 2007
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Zebrafish Tissue sections (T/S whole body. Spinal cord view)
Specification	T/S whole body. Spinal cord view
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: 10mM Citric acid pH6
Blocking step:	BSA as blocking agent for  10 mins  · Concentration: 1 % · Temperature: RT°C
Primary Antibody
Dilution	1/200
Incubation time:	16 hours  Diluent: TBS/BSA/Azide
Secondary Antibody
Name:	Non-Abcam Antibody was used: Goat anti mouse Igs Conjugation: Biotin
Dilution:	1/200
Additional Data
Rating	Poor
Additional notes	This Ab reagent is not applicable to zebra fish CNS/PNS, imho, for pwax zebrafish. I have tried dilution factors between 1/50-1/250. I attach a composite picture of this Ab and another polyclonal, from a different source, to support my statement. Upper image is "other source polyclonal", the lower image is Abcam's mouse monoclonal. NB: the adult zebrafish tissue was not decalcified, in case of loss of immunoreactivity. I used pressure-cooker HIER: thus bone tissue is mostly lost/disassociated.
Image	GFAP antibody [GF5] - Astrocyte Marker for IHC-P (Zebrafish) Copyright image used under license
Abcam response	Thank you for your feedback. It seems that this antibody previously untested in zebrafish is not suitable for this species. We hope this information will be useful to other researchers.

review by Carl Hobbs submitted on 19 November 2007
Application:	Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Rat Tissue sections (adult brain: lateral ventricle showing astrocytes)
Specification	adult brain: lateral ventricle showing astrocytes
Fixative:	Formaldehyde
Antigen retrieval step:	Heat mediated - Buffer/Enzyme Used: citric acid pH6
Permeabilization:	No
Blocking step:	BSA as blocking agent for  10 mins  · Concentration: 1 % · Temperature: RT°C
Primary Antibody
Dilution	1/250
Incubation time:	16 hours  Diluent: TBS/BSA/Azide pH7.6
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti mouse Ig Conjugation: Biotin
Dilution:	1/200
Additional Data
Rating	Average
Additional notes	In my hands this antibody reagent is not as sensitive as other anti -GFAP Abs that I use for P-Wax sections (a rabbit polyclonal and a mouse monoclonal Ab against GFAP). Picture shows the exit point for the progenitor olfactory neurones, of the lateral ventrical subventricular zone.
Image	GFAP antibody [GF5] - Astrocyte Marker for IHC-P (Rat) Copyright image used under license

review by Alexis Bemelmans submitted on 31 August 2007
Application:	Immunohistochemistry (Frozen sections)
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Rat Tissue sections (retina)
Specification	retina
Fixative:	Paraformaldehyde
Permeabilization:	Yes - 0.25% TritonX-100
Blocking step:	Serum as blocking agent for  1 hour  · Concentration: 10 % · Temperature: 20°C
Primary Antibody
Dilution	1/1000
Incubation time:	16 hours  Temperature: 20°C Diluent: PBS
Secondary Antibody
Name:	Non-Abcam Antibody was used: Anti Mouse IgG Conjugation: Alexa Fluor® 488
Dilution:	1/500
Additional Data
Rating	Excellent
Additional notes	Arrows indicate astrocytes processes at the inner side of the retina. Arrowhead indicates a Müller cell. Nuclei are counterstained in blue by Hoechst 33342. GCL: ganglion cell layer INL: inner nuclear layer.
Image	GFAP antibody [GF5] - Astrocyte Marker for IHC-Fr (Rat) Copyright image used under license

review by Randal Moldrich submitted on 16 December 2005
Application:	Immunocytochemistry
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Mouse Cell (mixed neuron/glia cell culture)
Specification	mixed neuron/glia cell culture
Fixative:	Paraformaldehyde
Blocking step:	BSA as blocking agent for  40 mins  · Concentration: 5%
Primary Antibody
Dilution	1/100
Incubation time:	12 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG Alexa Fluor 568 Conjugation:
Dilution:	1/400
Additional Data
Rating	Excellent
Additional notes	Image below: anti-GFAP[GF5] (ab10062) marked with red secondary antibody and TO-PRO-3 nuclear marker (660nm; converted to blue).
Image	GFAP antibody [GF5] - Astrocyte Marker for ICC (Mouse) Copyright image used under license

review by Randal Moldrich submitted on 16 December 2005
Application:	Immunocytochemistry
GFAP antibody [GF5] - Astrocyte Marker (ab10062)
Sample	Rat Cell (Glial cell culture)
Specification	Glial cell culture
Fixative:	Paraformaldehyde
Blocking step:	BSA as blocking agent for  40 mins  · Concentration: 5%
Primary Antibody
Dilution	1/100
Incubation time:	12 hours
Secondary Antibody
Name:	Non-Abcam Antibody was used: goat anti-mouse IgG Alexa Fluor 568 Conjugation:
Dilution:	1/400
Additional Data
Rating	Average
Additional notes	Image below: anti-GFAP (GF5) (ab10062) marked by a red secondary antibody with the nucleus counterstained with TO-PRO-3 (660nm, converted here to blue).
Image	GFAP antibody [GF5] - Astrocyte Marker for ICC (Rat) Copyright image used under license