Overview

  • Product name
    Anti-GFAP antibody [GF5]
    See all GFAP primary antibodies
  • Description
    Mouse monoclonal [GF5] to GFAP
  • 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, 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 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.

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 1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 43-45 kDa.
ELISA Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 20708681
IHC-Fr 1/1000.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC 1/100.

Target

  • Function
    GFAP, 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 specificity
    Expressed in cells lacking fibronectin.
  • Involvement in disease
    Defects 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 similarities
    Belongs to the intermediate filament family.
  • Post-translational
    modifications
    Phosphorylated by PKN1.
  • Cellular localization
    Cytoplasm. Associated with intermediate filaments.
  • Information by UniProt
  • Database links
  • Alternative names
    • wu:fb34h11 antibody
    • ALXDRD antibody
    • cb345 antibody
    • etID36982.3 antibody
    • FLJ42474 antibody
    • FLJ45472 antibody
    • Gfap antibody
    • GFAP_HUMAN antibody
    • gfapl antibody
    • Glial fibrillary acidic protein antibody
    • Intermediate filament protein antibody
    • wu:fk42c12 antibody
    • xx:af506734 antibody
    • zgc:110485 antibody
    see all

Anti-GFAP antibody [GF5] images

  • 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.

  • 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.

     

  • 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.

  • 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) 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.

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

This product has been referenced in:
  • Jongbloets BC  et al. Stage-specific functions of Semaphorin7A during adult hippocampal neurogenesis rely on distinct receptors. Nat Commun 8:14666 (2017). IF ; Mouse . Read more (PubMed: 28281529) »
  • Sun Y  et al. Therapeutic effect of apocynin through antioxidant activity and suppression of apoptosis and inflammation after spinal cord injury. Exp Ther Med 13:952-960 (2017). IHC ; Rat . Read more (PubMed: 28450925) »

See all 32 Publications for this product

Product Wall

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (spinal cord cross section, 25 micrometer thickness)
Permeabilization
No
Specification
spinal cord cross section, 25 micrometer thickness
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Formaldehyde
Username

Mr. Tamas Bellak

Verified customer

Submitted Nov 25 2015

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

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 colleagues in the lab to check whether they have experience using some of the antibodies ava...

Read More
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

Merci de votre intérêt pour ab10062.

Le laboratoire n'a malheureusement pas pu me dire si l'anticorps était spécifique à la forme soluble ou si il reconnaissait les deux formes, soluble et filamenteuse, de la GFAP. Ils ont pour l'instant toujo...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (differentiated cell from hES)
Specification
differentiated cell from hES
Fixative
Formaldehyde
Permeabilization
Yes - 0.25% Triton-X 100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 26°C
Username

Abcam user community

Verified customer

Submitted Sep 28 2012

Thank you for contacting us.

After completing a short literature search, GFAP is commonly used as an protoplasmic astrocyte marker. Below are two references:

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.00382...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line U87MG)
Specification
human glioblastoma cell line U87MG
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 06 2012

1-10 of 24 Abreviews or Q&A

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