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    gfap-antibody-ab7260.pdf

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Neuroscience Cell Adhesion Proteins Cytoskeletal Proteins Intermediate Filaments
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Anti-GFAP antibody (ab7260)

  • Datasheet
  • SDS
Reviews (97)Q&A (10)References (557)

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Immunohistochemistry (PFA fixed) - Anti-GFAP antibody (ab7260)
  • Western blot - Anti-GFAP antibody (ab7260)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
  • Western blot - Anti-GFAP antibody (ab7260)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
  • Immunocytochemistry - Anti-GFAP antibody (ab7260)

Key features and details

  • Rabbit polyclonal to GFAP
  • Suitable for: IHC (PFA fixed), WB, IHC-P, ICC
  • Reacts with: Mouse, Rat
  • Isotype: IgG

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Overview

  • Product name

    Anti-GFAP antibody
    See all GFAP primary antibodies
  • Description

    Rabbit polyclonal to GFAP
  • Host species

    Rabbit
  • Specificity

    Specifically 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 applications

    Suitable for: IHC (PFA fixed), WB, IHC-P, ICCmore details
  • Species reactivity

    Reacts with: Mouse, Rat
    Predicted to work with: Horse, Cow, Human, Pig, Mammals
  • Immunogen

    Recombinant full length protein corresponding to Human GFAP. Isotype 1 expressed in and purified from E. coli.

  • General notes

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
  • Concentration information loading...
  • Purity

    Whole antiserum
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Adhesion Proteins
    • Cytoskeletal Proteins
    • Intermediate Filaments
    • Neuroscience
    • Cell Type Marker
    • Glia marker
    • Astrocyte marker
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class II
    • GFAP
    • Tags & Cell Markers
    • Cell Type Markers
    • Neuroscience Markers
    • Glial
    • Stem Cells
    • Neural Stem Cells
    • Glial Restricted Lineage
    • Astrocyte

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human GFAP protein (ab114149)
    • Recombinant Human GFAP protein (ab151370)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-GFAP antibody [ZRF-1] (ab154474)
    • Human GFAP Matched Antibody Pair Kit (ab222279)

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 Abreviews Notes
IHC (PFA fixed) Use at an assay dependent concentration.
WB 1/10000. Detects a band of approximately 55,48 kDa.

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

IHC-P 1/1000 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/5000.

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.
  • 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

    • Entrez Gene: 281189 Cow
    • Entrez Gene: 2670 Human
    • Entrez Gene: 14580 Mouse
    • Entrez Gene: 396562 Pig
    • Entrez Gene: 24387 Rat
    • Omim: 137780 Human
    • SwissProt: Q28115 Cow
    • SwissProt: P14136 Human
    • SwissProt: P03995 Mouse
    • SwissProt: P47819 Rat
    • Unigene: 514227 Human
    • Unigene: 1239 Mouse
    • Unigene: 91512 Rat
    see all
  • 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

  • Immunohistochemistry (PFA fixed) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (PFA fixed) - Anti-GFAP antibody (ab7260)

    GFAP antibody ab7260 was used with Tissue Clearing Kit ab243298 to penetrate, stain and clear a 1 mm coronal section of mouse brain. Blue: DAPI, Green: GFAP.

    Learn more about tissue clearing kits, reagents, and protocols designed to make it easier to stain thick tissue sections and get more data from each valuable tissue section.

    To use this antibody with tissue clearing, use Tissue Clearing Kit ab243298. For 1 mm brain sections, we recommend a starting dilution of 1:1000, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1:400.

  • Western blot - Anti-GFAP antibody (ab7260)
    Western blot - Anti-GFAP antibody (ab7260)This image is courtesy of an anonymous Abreview
    All lanes : Anti-GFAP antibody (ab7260) at 1/5000 dilution

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

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : 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

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)

    IHC image of GFAP staining in a formalin fixed, paraffin embedded normal mouse brain tissue section, performed on a Leica Bond™ system using the standard protocol B.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7260 at 1/2000 dilution for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with hematoxylin 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)Yoon, H. et al PLoS One. 2017 Jul 10;12(7):e0180697. doi: 10.1371/journal.pone.0180697. eCollection 2017 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Increases in GFAP after demyelinating injury are greater in the spinal cord compared to brain

    Photomicrographs show immunoreactivity for GFAP or ALDH1L1 in (A) the corpus callosum, or (B) the dorsal column white matter of adult mice at base line, and at 14 d after microinjection of the demyelinating agent lysolecithin. Histograms show the percent area of GFAP immunofluorescence, and expression of GFAP/ALDH1L1+ astrocyte, was significantly greater in the spinal cord compared to the corpus callosum 14d post-lysolecithin lesion.

    GFAP was detected using ab7260.

    (From Figure 4A and 4B of Hoon et al)

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

    Lane 1 : Rat brain lysate
    Lane 2 : Rat spinal cord lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Mouse spinal cord lysate
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)

    IHC image of GFAP staining in a formalin fixed, paraffin embedded normal rat 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 (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7260 at 1/2000 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 hematoxylin 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.

  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
    Immunocytochemistry - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Chia-Li Lin.

    ab7260 staining GFAP in cells from mouse brain tissue sections by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 in PBS and blocked with 1% BSA for 40 minutes at 25°C. Samples were incubated with primary antibody (1/1200 in TBS) for 24 hours at 4°C. Goat Anti-Rabbit IgG H&L (DyLight® 488) (ab96883) was used as the secondary antibody at a dilution of 1/200.

    See Abreview

  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
    Immunocytochemistry - Anti-GFAP antibody (ab7260)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Mr Osama Mohsen

    ab7260 staining rat brain tissue sections by IHC-P.

    Sections were fixed in formaldehyde and blocked with a commercialy available blocking agent prior to incubating with ab7260, diluted 1/5000 for 20 hours at 4°C.  An HRP conjugated mouse polyclonal (universal HRP polymer detection) antibody was used as the secondary.

    See Abreview

  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
    Immunocytochemistry - Anti-GFAP antibody (ab7260)This image is courtesy of an Abreview submitted by Ms Nancy Nutile-McMenemy

     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 visualized using DAPI.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody (ab7260)This image was kindly supplied by Dr Vladimir Milenkovic by 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

  • Immunocytochemistry - Anti-GFAP antibody (ab7260)
    Immunocytochemistry - Anti-GFAP antibody (ab7260)

    GFAP antibody ab7260 was used with 3D Cell Culture Clearing Kit ab243299 to penetrate, stain and clear a 3D neuronal spheroid cell culture. Blue: DAPI, Green: GFAP.

    Learn more about 3D cell culture clearing and tissue clearing kits, reagents, and protocols designed to make it easier to stain 3D cell cultures and thick tissue sections and get more data from each valuable tissue section. 

    To use this antibody with clearing, use our 3D Cell Culture Clearing Kit ab243299. We recommend a starting dilution of 1:1000, and also using Goat Anti-Rabbit IgG H&L AlexaFluor488 (ab150077) at a dilution of 1:400.

Protocols

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (557)

    Publishing research using ab7260? Please let us know so that we can cite the reference in this datasheet.

    ab7260 has been referenced in 557 publications.

    • Findlay AS  et al. Fam151b, the mouse homologue of C.elegans menorin gene, is essential for retinal function. Sci Rep 10:437 (2020). PubMed: 31949211
    • Yin N  et al. IDH1-R132H mutation radiosensitizes U87MG glioma cells via epigenetic downregulation of TIGAR. Oncol Lett 19:1322-1330 (2020). PubMed: 31966064
    • Tu Y  et al. Melatonin inhibits Müller cell activation and pro-inflammatory cytokine production via upregulating the MEG3/miR-204/Sirt1 axis in experimental diabetic retinopathy. J Cell Physiol N/A:N/A (2020). PubMed: 32324260
    • Li W  et al. Dysfunctional Nurr1 promotes high glucose-induced Müller cell activation by up-regulating the NF-?B/NLRP3 inflammasome axis. Neuropeptides N/A:102057 (2020). PubMed: 32461025
    • Zhao M  et al. Vinpocetine Protects Against Cerebral Ischemia-Reperfusion Injury by Targeting Astrocytic Connexin43 via the PI3K/AKT Signaling Pathway. Front Neurosci 14:223 (2020). PubMed: 32300287
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (10 micon thick spinal cord sections)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Vector Laboratories Cat# H-3300
    Permeabilization
    Yes - 0.5% Triton
    Specification
    10 micon thick spinal cord sections
    Blocking step
    normal donkey serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 09 2020

    Immunohistochemistry (Frozen sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - 0.05% triton-x100
    Specification
    Brain
    Blocking step
    Vetor Blocker as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 07 2019

    Western blot abreview for Anti-GFAP antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Brain)
    Gel Running Conditions
    Reduced Denaturing (4-12% gel)
    Loading amount
    20 µg
    Specification
    Brain
    Blocking step
    Protein Free blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 18 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (spinal cord)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
    Permeabilization
    No
    Specification
    spinal cord
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 09 2019

    Immunohistochemistry free floating abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Adult rat brainstem tissue)
    Specification
    Adult rat brainstem tissue
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 28 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain cortex)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA
    Permeabilization
    No
    Specification
    Brain cortex
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 20 2018

    Immunohistochemistry free floating abreview for Anti-GFAP antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Brain)
    Specification
    Brain
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 30 2018

    Western blot abreview for Anti-GFAP antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (10%)
    Loading amount
    30 µg
    Treatment
    250 μg/kg LPS for 1 week
    Specification
    Brain
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Ms. Ji Hye Han

    Verified customer

    Submitted Jul 31 2018

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (rat spinal cord)
    Antigen retrieval step
    None
    Permeabilization
    Yes - PBS + 0.2% triton + 4% BSA
    Specification
    rat spinal cord
    Blocking step
    BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Alexandre Magno

    Verified customer

    Submitted Jul 20 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-GFAP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Ms. Ji Hye Han

    Verified customer

    Submitted May 24 2018

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