Overview

  • Product nameAnti-GFAP antibodySee all GFAP primary antibodies ...
  • Description
    Rabbit polyclonal to GFAP
  • SpecificitySpecifically recognizes mammalian GFAP on western blots and immunocytochemically. Detects a band of 55kDa corresponding to GFAP and also a GFAP derived 48kDa band. Some customers have successfully used ab7260 on Zebrafish lysates; however we have conflicting data to suggest that not all batches will be suitable for work on Zebrafish. For further information, please contact Abcam Scientific Support.
  • Tested applicationsIHC-FoFr, IHC-Fr, ICC/IF, WB, IHC-P, ICC more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cat, Human, Marmoset (common)
    Predicted to work with: Cow, Pig, all Mammals
  • Immunogen

    The initial immunization was performed with a preparation of full length human recombinant GFAP expressed in bacteria and highly purified. Subsequent boosts were performed with GFAP purified from a Triton X-100 extract of myelin associated material from bovine spinal cord, following an "axonal flotation" procedure (Liem et al.). The GFAP was further purified by centrifugation and ion exchange chromatography in 6m urea on DEAE cellulose.

  • General notesIn some cases, the antibody may appear red in color. This is due to small amounts of hemolysis, and does not affect antibody performance.

Properties

Applications

Our Abpromise guarantee covers the use of ab7260 in the following tested applications.

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

Application Notes
IHC-FoFr 1/5000. See Abreview.
IHC-Fr 1/500.
ICC/IF 1/1000.
WB 1/50000. Detects a band of approximately 55,48 kDa.

This lower 48kDa band is thought to be a degradation product.

IHC-P 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC 1/5000.

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.
  • Target information above from: UniProt accession P14136 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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 images

  • Immunohistochemistical detection of GFAP antibody - Astrocyte Marker (ab7260) on formaldehyde-fixed paraffin-embedded monkey brain sections. Antigen retrieval step: Heat mediated.  Buffer Used: Citric acid pH6. Permeabilization: None. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody incubated at 1/2000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti rabbit IgG Conjugated to biotin  (1/200). Marmoset brain: astrocytes are clearly and strongly labelled.

    See Abreview

  • ab7260 at 1/10000 dilution staining Mouse cortical astrocytes by Immunocytochemistry. The cells were permeabilized with Triton/HEPES buffer prior to primary application. The antibody was incubated with the cells for 18 hours and then bound antibody was detected with an Alexa Fluor ® 488 conjugated goat anti-rabbit antibody.

    This image is courtesy of an Abreview submited by Charmaine Noonan.

    See Abreview

  •  ab7260 at a 1/5000 dilution staining rat spinal cord tissue sections by IHC-Fr. Rats were transcardially perfused with 4% PFA. The tissue was post fixed 1 hour in 4% PFA and then 30% sucrose for three days. 20µm sections were cryostat cut. The primary antibody was incubated with the tissue sections for 18 hours. Bound antibody was detected using an Alexa Fluor ® 488 conjugated goat anti-rabbit polyclonal.

    See Abreview

  • Western blot of whole rat cerebellum homogenate stained with ab7260 at dilution of 1:100,000. A prominent band running with an apparent SDS-PAGE molecular weight of ~55kDa corresponds to rodent GFAP. A lower band at ~48kDa is derived from the GFAP molecule.

  • ab7260 staining rat brain tissue sections by IHC-P.  Sections were fixed in formaldehyde and bocoked with a commercialy available blocking agent prior to incubating with ab7260, diluted 1/5000 for 20 hours at 4°C.  A HRP conjugated mouse polyclonal (universal HRP polymer detection) antibody was used as the secondary.

    See Abreview

  • All lanes : Anti-GFAP antibody (ab7260) at 1/5000 dilution

    Lane 1 : Rat thoracotomy, spinal cord homogenate
    Lane 2 : Rat thoracotomy, spinal cord homogenate
    Lane 3 : Rat thoracotomy, spinal cord homogenate
    Lane 4 : Rat thoracotomy sham, spinal cord homogenate
    Lane 5 : Rat thoracotomy sham, spinal cord homogenate
    Lane 6 : Rat nerve transect sham, spinal cord homogenate
    Lane 7 : Rat nerve transect sham, spinal cord homogenate
    Lane 8 : Rat nerve transect, spinal cord homogenate
    Lane 9 : Rat nerve transect, spinal cord homogenate

    Lysates/proteins at 30 µg per lane.

    Secondary
    HRP conjugated goat anti-rabbit at 1/3000 dilution
    developed using the ECL technique

    Observed band size : 53 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab7260 staining GFAP in mouse eye tissue sections by Immunohistochemistry (paraffin embedded sections). Tissue was fixed with paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then permeabilized using 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C; followed by incubation with the primary antibody, at a 1/500 dilution, for 16 hours at 4°C. The secondary antibody used was a goat anti-rabbit IgG conjugated to Alexa Fluor® 488 used at a 1/5000 dilution.
    The retinal layers are: ganglion cells layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), and photoreceptor outer segments (ROS). Nuclei were counterstained with DAPI.

    See Abreview

  •  ab7260 staining rat pup cortical preps by ICC/IF. The preps were grown for 14 days in culture and plated onto coverslips. The preps were acid/alcohol fixed and blocked prior to incubation with ab7260. Bound antibody was detected using an Alexa Fluor ®488 conjugated goat polyclonal antibody. Nuclei were visualised using DAPI.

    See Abreview

References for Anti-GFAP antibody (ab7260)

This product has been referenced in:
  • Bettum IJ  et al. Metastasis-associated protein S100A4 induces a network of inflammatory cytokines that activate stromal cells to acquire pro-tumorigenic properties. Cancer Lett 344:28-39 (2014). Read more (PubMed: 24215866) »
  • Smith AJ  et al. Improving lithium therapeutics by crystal engineering of novel ionic cocrystals. Mol Pharm 10:4728-38 (2013). ICC/IF ; Rat . Read more (PubMed: 24191685) »

See all 25 Publications for this product

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0 with 0.05% Tween 20
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization No
Fixative Paraformaldehyde
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Submitted Sep 24 2013

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (12%)
Sample Mouse Cell lysate - whole cell (Neuro2A- neuroblastoma)
Specification Neuro2A- neuroblastoma
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Jul 01 2013

Abreviews
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HNGC-2)
Specification HNGC-2
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Jun 04 2013

Abreviews
Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Human Cultured Cells (U87 Human primary Glioblastoma cell line)
Specification U87 Human primary Glioblastoma cell line
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
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Submitted Jun 04 2013

Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (nerve)
Specification nerve
Fixative Paraformaldehyde
Permeabilization Yes - 0.2% triton
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted May 08 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Mouse, brain tissue, whole brain sections)
Specification Mouse, brain tissue, whole brain sections
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - Tween-20
Blocking step Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Submitted Apr 16 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...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (Glioblastoma primary cells)
Loading amount 30 µg
Specification Glioblastoma primary cells
Gel Running Conditions Reduced Denaturing (gradient gel 5-15%)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
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Submitted Dec 12 2012

Thank you for contacting Abcam.

When comparing multiple products that have been tested in the same species and applications, we like to use the abreviews and specific references to judge the potential for an antibody to perform as indicated on...

Read More
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Neural culture)
Specification Neural culture
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 21 2012

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