Overview

  • Product nameAnti-GFAP antibody [GF5]See all GFAP primary antibodies ...
  • Description
    Mouse monoclonal [GF5] to GFAP
  • SpecificityThere is no cross-reactivity with other neurospecific proteins.
  • Tested applicationsIHC-P, WB, ELISA, IHC-FoFr, IHC-Fr, ICC/IF, Flow Cyt, ICCmore details
  • 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
  • General notesConcentration varies from lot to lot and can be provided on request.

Properties

Applications

Our Abpromise guarantee covers the use of ab10062 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/250 - 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent dilution. Detects a band of approximately 43-45 kDa.
ELISA Use at an assay dependent dilution.
IHC-FoFr Use at an assay dependent dilution. PubMed: 20708681
IHC-Fr 1/1000.
ICC/IF Use at an assay dependent dilution.
Flow Cyt Use 1-2µg for 106 cells.
ICC 1/100.

Target

  • FunctionGFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells.
  • Tissue specificityExpressed in cells lacking fibronectin.
  • Involvement in diseaseDefects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Post-translational
    modifications
    Phosphorylated by PKN1.
  • Cellular localizationCytoplasm. Associated with intermediate filaments.
  • Information by UniProt
  • Database links
  • Alternative names
    • Astrocyte antibody
    • FLJ42474 antibody
    • FLJ45472 antibody
    • GFAP antibody
    • GFAP_HUMAN antibody
    • Glial Fibrillary Acidic Protein antibody
    • Intermediate filament protein antibody
    see all

Anti-GFAP antibody [GF5] images

  • 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.

  • 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.

  • 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.
  • 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

  • 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.

References for Anti-GFAP antibody [GF5] (ab10062)

This product has been referenced in:
  • Lang B  et al. Recurrent deletions of ULK4 in schizophrenia: a gene crucial for neuritogenesis and neuronal motility. J Cell Sci 127:630-40 (2014). Read more (PubMed: 24284070) »
  • Searcy JL  et al. Impact of age on the cerebrovascular proteomes of wild-type and Tg-SwDI mice. PLoS One 9:e89970 (2014). WB ; Mouse . Read more (PubMed: 24587158) »

See all 14 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Mouse Tissue sections (frozen brain section)
Specification frozen brain section
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 23 2013

Application Flow Cytometry
Fixation Frozen
Permeabilization Yes - 0.1% Triton-X
Sample Human Cell (Brain)
Specification Brain
Gating Strategy forward scatter/side scatter (all cells)
Preparation Cell harvesting/tissue preparation method: Homogenized frozen tissue
Sample buffer: Tris/EDTA/sucrose/Triton
Username

Dr. Mollie Woodworth

Verified customer

Submitted Sep 03 2013

Thank you for contacting us. I apologise for the delay getting back to you.

Several markers can be used to stain astrocytes: alpha Smooth Muscle Actin, Bystin, GFAP, GLT-1, GLAST, S100, Mab 6.17, SC1, etc…

I have contacted my...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - 0.03% Trition
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Astrocytes)
Loading amount 30 µg
Specification Astrocytes
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Feb 08 2013

Thank you for getting back to me.

I could certainly arrange the new vial (ab7260) as a replacement for you as requested.

However, after reading through all the previous correspondence, I would like to highlight and advise the follow...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for getting back to me. I am very sorry to hear that the product does not perform.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means th...

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Thank you for your enquiry regarding ab10062 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

This antibody has not been tested specifically in...

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I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application us...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"