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    products/primary-antibodies/gfap-antibody-epr1034y-ab68428.pdf

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Neuroscience Cell Adhesion Proteins Cytoskeletal Proteins Intermediate Filaments
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RecombinantRabMAb

Recombinant Anti-GFAP antibody [EPR1034Y] (ab68428)

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Reviews (11)Q&A (1)References (45)

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Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
  • Anti-GFAP antibody [EPR1034Y] (ab68428)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1034Y] to GFAP - N-terminal
  • Suitable for: WB, IP, IHC-P, ICC/IF, mIHC, Flow Cyt (Intra), IHC-Fr
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-GFAP antibody [EPR1034Y]
    See all GFAP primary antibodies
  • Description

    Rabbit monoclonal [EPR1034Y] to GFAP - N-terminal
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IF, mIHC, Flow Cyt (Intra), IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • mIHC: Human cerebrum tissue and human cerebellum tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.21% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR1034Y
  • 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
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-GFAP antibody [EPR1034Y] (ab194324)
    • Alexa Fluor® 647 Anti-GFAP antibody [EPR1034Y] (ab194325)
    • Alexa Fluor® 594 Anti-GFAP antibody [EPR1034Y] (ab201732)
    • Alexa Fluor® 555 Anti-GFAP antibody [EPR1034Y] (ab201735)
    • Alexa Fluor® 568 Anti-GFAP antibody [EPR1034Y] (ab201736)
    • Alexa Fluor® 405 Anti-GFAP antibody [EPR1034Y] (ab206586)
    • Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)
    • Anti-GFAP antibody [EPR1034Y] (N-terminal) - Chimeric (ab302644)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

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

    • Human GFAP Matched Antibody Pair Kit (ab222279)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68428 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
WB
1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

For unpurified use at 1/50 000 - 1/100 000.

IP
1/20 - 1/40.
IHC-P (6)
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF (1)
Use a concentration of 0.1 - 1 µg/ml.
mIHC
Use at an assay dependent concentration.
Flow Cyt (Intra)
1/500.
IHC-Fr (2)
1/250.
Notes
WB
1/10000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).

For unpurified use at 1/50 000 - 1/100 000.

IP
1/20 - 1/40.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF
Use a concentration of 0.1 - 1 µg/ml.
mIHC
Use at an assay dependent concentration.
Flow Cyt (Intra)
1/500.
IHC-Fr
1/250.

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: 2670 Human
    • Entrez Gene: 14580 Mouse
    • Entrez Gene: 24387 Rat
    • Omim: 137780 Human
    • 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

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Fluorescence multiplex immunohistochemical analysis of the human cerebellum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-P2Y12 (ab254347 red; Opal™570) on human cerebellum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and ab254347 (1/1000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Fluorescence multiplex immunohistochemical analysis of the human cerebrum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-P2Y12 (ab254347, red; Opal™570) on human cerebrum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and ab254347 (1/1000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary brain cells cells labelling GFAP with ab68428 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    IHC image of GFAP staining in a formalin fixed, paraffin embedded normal human 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 ab68428 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunofluorescence staining of GFAP using ab68428 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab68428 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab68428 gave comparable results using 4% formaldehyde fixation (10 min).

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Fluorescence multiplex immunohistochemical analysis of human cerebrum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-MAP2 (ab254263, red; Opal™570) on human cerebrum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and ab254263 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/10000 dilution

    Lane 1 : Human cerebellum tissue lysate at 20 µg
    Lane 2 : Human brain tissue lysate at 10 µg

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 50 kDa
    Observed band size: 48-50 kDa why is the actual band size different from the predicted?



    Blocking and Diluting buffer 5% NFDM/TBST
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural mix culture cells labelling GFAP with ab68428 at 1/250 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab10062 anti-GFAP antibody [GF5] at 1/100 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2µg/ml) was used as a counterstain. The Nuclear counterstain was DAPI (Blue).

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling GFAP with ab68428 at 1/1000 dilution (2.011 µg/mL) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).<\p>

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (ab68428, gray; Opal™690), anti-Myelin PLP (ab254363, green; Opal™520) and anti-MAP2 (ab254263, red; Opal™570) on human cerebellum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of ab68428 (1/50 dilution), ab254363 (1/2000 dilution), and ab254263 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin-fixed paraffin-embedded section).

    Merged staining of Neu-N (ab177487; yellow; Opal™570), anti-beta III Tubulin (ab52623; red; Opal™690) and anti-GFAP (ab68428; green; Opal™520).

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ kit.

    The section was incubated in three rounds of staining with ab177487 (1/1000 dilution), ab52623 (1/200 dilution) and ab68428 (1/250 dilution); each using a separate fluorescent tyramide signal amplification system.

    Sodium citrate antigen retrieval (pH 6.0) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

    DAPI (blue) was used as a nuclear counter stain.

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Panel A: Merged staining of anti-GFAP (gray; Opal™690), anti-P2Y12 (green; Opal™520) and anti-MAP2 (red; Opal™570) on human cerebrum.
    Panel B: Anti-MAP2 stained cell body and dendrites of neurons.
    Panel C: Anti-P2Y12 stained on microglial cells.
    Panel D: Anti-GFAP stained on astrocytes.

    The section was incubated in three rounds of staining: in the order of ab68428, ab254347, and ab254263 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  • Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Panel A: Merged staining of anti-GFAP (gray; Opal™690), anti-P2Y12 (green; Opal™520) and anti-MAP2 (red; Opal™570) on human cerebellum.
    Panel B: Anti-MAP2 stained cell body and dendrites of neurons.
    Panel C: Anti-P2Y12 stained on microglial cells.
    Panel D: Anti-GFAP stained on astrocytes.

    The section was incubated in three rounds of staining: in the order of ab68428, ab254347, and ab254263 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling GFAP with ab68428 at 1/1000 dilution (2.011 µg/mL) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).<\p>

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    IHC image of GFAP staining in a section of frozen normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab68428, 1/250 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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/10000 dilution

    Lane 1 : Mouse cerebellum tissue lysate
    Lane 2 : Mouse brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 50 kDa
    Observed band size: 50 kDa



    Blocking and Diluting buffer 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/50000 dilution

    Lane 1 : Rat cerebellum tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 50 kDa
    Observed band size: 50 kDa



    Blocking and Diluting buffer 5% NFDM/TBST
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. 

  • Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (ab68428)

    ab68428 at 1/20 dilution immunoprecipitating GFAP in rat brain whole cell lysate observed at 50 KDa (lanes 1 and 2).

    Lane 1 (input): Rat brain whole cell lysate 10ug

    Lane 2 (+): ab68428 + Rat brain whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68428 in Rat brain whole cell lysate

    For western blotting, ab68428 was used followed by VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. 

    Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

  • Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Western blot - Anti-GFAP antibody [EPR1034Y] (ab68428)
    All lanes : Anti-GFAP antibody [EPR1034Y] (ab68428) at 1/5000 dilution (unpurified)

    Lane 1 : Human brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti-Rabbit antibody at 1/2000 dilution

    Predicted band size: 50 kDa
    Observed band size: 50 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of formalin-fixed paraffin-embedded human brain (left) and human glioma (right) tissue sections labelling GFAP with unpurified ab68428 at dilution of 1/250.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue sections labelling GFAP with purified ab68428 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] (ab68428)

    Immunohistochemical analysis of formalin-fixed paraffin-embedded mouse brain tissue section labelling GFAP with unpurified ab68428 at dilution of 1/250.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-GFAP antibody [EPR1034Y] (ab68428)
    Anti-GFAP antibody [EPR1034Y] (ab68428)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (45)

Publishing research using ab68428? Please let us know so that we can cite the reference in this datasheet.

ab68428 has been referenced in 45 publications.

  • Kim HM  et al. The epidermal growth factor receptor variant type III mutation frequently found in gliomas induces astrogenesis in human cerebral organoids. Cell Prolif 54:e12965 (2021). PubMed: 33283409
  • Zheng Y  et al. Spatial learning and memory deficits induced by prenatal glucocorticoid exposure depend on hippocampal CRHR1 and CXCL5 signaling in rats. J Neuroinflammation 18:85 (2021). PubMed: 33810797
  • Dang TM  et al. Optimization of the isolation procedure and culturing conditions for hepatic stellate cells obtained from mouse. Biosci Rep 41:N/A (2021). PubMed: 33350435
  • Button EB  et al. Development of a novel, sensitive translational immunoassay to detect plasma glial fibrillary acidic protein (GFAP) after murine traumatic brain injury. Alzheimers Res Ther 13:58 (2021). PubMed: 33678186
  • Schwabenland M  et al. Deep spatial profiling of human COVID-19 brains reveals neuroinflammation with distinct microanatomical microglia-T-cell interactions. Immunity 54:1594-1610.e11 (2021). PubMed: 34174183
View all Publications for this product

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Immunocytochemistry/ Immunofluorescence abreview for Anti-GFAP antibody [EPR1034Y]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Syrian hamster Cell (Brain tissue)
Permeabilization
Yes - 1% Triton X-100 in PBS
Specification
Brain tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Apr 21 2021

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