Overview

  • Product name
    Anti-GFAP antibody [GF5]
    See all GFAP primary antibodies
  • 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, 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

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.

  • 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.

References

This product has been referenced in:
  • Lu Y  et al. Fibroblast growth factor 21 facilitates peripheral nerve regeneration through suppressing oxidative damage and autophagic cell death. J Cell Mol Med 23:497-511 (2019). Read more (PubMed: 30450828) »
  • Yang D  et al. Normobaric oxygen inhibits AQP4 and NHE1 expression in experimental focal ischemic stroke. Int J Mol Med 43:1193-1202 (2019). Read more (PubMed: 30592266) »
See all 70 Publications for this product

Customer reviews and Q&As

21-27 of 27 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
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 minute(s) · Concentration: 1% · Temperature: RT°C

Mr. Carl Hobbs

Verified customer

Submitted Dec 04 2007

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
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 minute(s) · Concentration: 1% · Temperature: RT°C

Mr. Carl Hobbs

Verified customer

Submitted Nov 19 2007

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (retina)
Specification
retina
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% TritonX-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. Alexis Bemelmans

Verified customer

Submitted Aug 31 2007

Answer

Thank you for your enquiry. I am sorry it has taken some time to find some information for you. As far as I am aware, this antibody (ab10062) has been tested in ELISA but unfortunately, there is no data available on the minimal detectable limits or specificity.

Read More

Answer

The correct isotype control for ab10062 is ab18428, while the correct isotype control for ab18460 is ab18449.

Read More
Application
Immunocytochemistry
Sample
Rat Cell (Glial cell culture)
Specification
Glial cell culture
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5%

Dr. Randal Moldrich

Verified customer

Submitted Dec 16 2005

Application
Immunocytochemistry
Sample
Mouse Cell (mixed neuron/glia cell culture)
Specification
mixed neuron/glia cell culture
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5%

Dr. Randal Moldrich

Verified customer

Submitted Dec 16 2005

21-27 of 27 Abreviews or Q&A

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