Overview

  • Product name
  • Description
    Chicken polyclonal to GFAP
  • Host species
    Chicken
  • Tested applications
    Suitable for: ELISA, IHC-Fr, IHC-FoFr, IHC-P, Flow Cyt, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Human, Rhesus monkey, Apteronotus leptorhynchus
    Predicted to work with: Mammals
  • Immunogen

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

  • Positive control
    • ICC/IF: Rat astrocytes. WB: Rat and mouse brain lysate. Apteronotus leptorhynchus brain tissue lysate. IHC-P: Rat hippocampus tissue. Mouse brain tissue. IHC-Fr: Human brain tissue. IHC-PFA fixed frozen: Mouse hippocampal tissue. Rabbit eye tissue. Flow Cyt: Human brain cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab4674 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
ELISA 1/5000.
IHC-Fr 1/1000 - 1/5000. Try this antibody at about between about 1:1,000 using fluorescent secondary antibodies or 1:5,000 using peroxidase or other enzyme linked methods.
IHC-FoFr 1/1000 - 1/5000. PubMed: 20098733Try this antibody at about between about 1:1,000 using fluorescent secondary antibodies or 1:5,000 using peroxidase or other enzyme linked methods.
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/100.

ab37382 - Chicken polyclonal IgY, is suitable for use as an isotype control with this antibody.

We recommend using ab46969 goat anti-chicken IgY FITC secondary antibody.

WB 1/1000 - 1/5000. Predicted molecular weight: 50 kDa.

Expect to see a band at 55kDa and another at about 48kDa, apparently a breakdown product of the 55kDa band.

ICC/IF 1/1000 - 1/5000.

Try this antibody at about between about 1:1,000 using fluorescent secondary antibodies or 1:5,000 using peroxidase or other enzyme linked methods.

PubMed: 25418722

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

  • Rat astrocytes stained with ab4674 (red). The anti-GFAP antibody was used at a dilution of 1:50 from affinity purified material using our standard fixation and staining procedure (in protocol section). Hoechst dye reveals nuclear DNA in blue.
  • Immunofluorescent analysis of a section of mouse hippocampus stained with ab4674 at a 1:5,000 dilution in green.

    Costained with a rabbit pAb to FOX3/NeuN dilution 1:5,000, in red. The blue is DAPI staining of nuclear DNA. Following transcardial perfusion with 4% paraformaldehyde, mouse brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections were stained. The GFAP antibody stains a network of astroglial cells while the Fox3/NeuN antibody stains the nuclei and proximal perikarya of neurons.

  • IHC image of GFAP staining in a formalin-fixed, paraffin-embedded mouse normal brain tissue section.

    The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab4674 at 1/1000 dilution for 15 minutes at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

  • All lanes : Anti-GFAP antibody (ab4674) at 1/5000 dilution

    Lane 1 : Rat whole brain lysate
    Lane 2 : Mouse whole brain lysate

    Predicted band size: 50 kDa

  • ab4674 staining GFAP in mouse hippocampus tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Tissue samples were fixed by perfusion with 4% PFA and blocked with 10% serum for 30 minutes at 25°C. The sample was incubated with primary antibody (1/500) for 16 hours at 25°C in 10% NGS in PBS + 0.1% TX100. An Alexa Fluor®488-conjugated Goat polyclonal to chicken IgG was used as secondary antibody at 1/400 dilution. Staining was intensified with 2-3 minutes of retrieval with trypsin at room temperature.

    See Abreview

  • ab4674 staining GFAP in human brain cells by Flow Cytometry.

    Cells were grown in stem cell media, RHB-A, and collected using 0.05% trypsin + trypsin inhibitor. Cells were fixed in 80% Acetone for 5-10 minutes at -20°C. The sample was incubated with the primary antibody (1/100 in 10% FCS in PBS) for 20 minutes at 25°C. ab46969 a goat anti-chicken IgY FITC (1/100) was used as the secondary antibody
    Gating Strategy: Cells expressing GFAP

    See Abreview

  • ab4674 staining GFAP in rat primary astrocytes by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with paraformaldehyde, permeabilized with 0.05% Triton X-100 and blocked with 5% serum for 20 minutes at 20°C. Samples were incubated with primary antibody (1/2000) for 24 hours at 4°C. ab6873, a goat anti-chicken IgY FITC (1/1000) was used as the secondary antibody.

    See Abreview

  • Frozen sectioned human brain stained for GFAP with ab4674 (1/500 dilution, 18 hours at 4°C) in immunohistochemical analysis.

    10 µm cryostat cut sections fixed with 4% PFA prior to staining for 10 minutes. Goat polyclonal anti-chicken-Alex-Fluor®647 was used as the secondary at a 1/250 dilution.

    Micrograph demonstrates astrocyte processes (blue) next to cerebral vessels (red).

    See Abreview

  • ab4674 staining GFAP in rabbit eye tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Tissue samples were fixed by perfusion with formaldehyde and blocked with BSA for 2 hours at 4°C. The sample was incubated with primary antibody (1/1000) at 4°C for 12 hours. ab150175, a goat anti-chicken IgY Alexa Fluor® 647 (1/1000), was used as the secondary antibody.

    See Abreview

  • Anti-GFAP antibody (ab4674) at 1/1000 dilution + Apteronotus leptorhynchus brain tissue lysate at 50 µg

    Secondary
    AlexaFluor®488-conjugated goat anti-chicken polyclonal IgG at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 5 minutes

    See Abreview

  • IHC image of GFAP staining in a formalin fixed, paraffin embedded normal rat hippocampus tissue section.

    The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab4674 at 1/1000 dilution for 15 minutes at room temperature. A goat anti-chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

  • ab4674 detecting recombinant GFAP by direct ELISA.

    Mouse recombinant GFAP protein was coated onto a microplate in carbonate coating buffer pH 9.6 for 1 hour at 37°C. Plate was blocked with 3% BSA for 1 hour at 37°C and incubated with the primary antibody (1/5000 in PBS + 1% Tween-20) for 1 hour at 37°C. An undiluted alkaline phosphatase Goat anti-chicken IgG polyclonal was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Stark DT  et al. Optic Nerve Regeneration After Crush Remodels the Injury Site: Molecular Insights From Imaging Mass Spectrometry. Invest Ophthalmol Vis Sci 59:212-222 (2018). Read more (PubMed: 29340649) »
  • Lam LKM  et al. Gamma-interferon exerts a critical early restriction on replication and dissemination of yellow fever virus vaccine strain 17D-204. NPJ Vaccines 3:5 (2018). Read more (PubMed: 29387474) »

See all 134 Publications for this product

Customer reviews and Q&As

Application
IHC - Wholemount
Sample
Mouse Tissue (Brain)
Specification
Brain
Username

Mr. Gabriel Luna

Verified customer

Submitted Jan 05 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 4°C
Antigen retrieval step
None
Sample
Rabbit Tissue sections (eye)
Specification
eye
Permeabilization
No
Fixative
Formaldehyde
Username

Dr. Alex Zagariya

Verified customer

Submitted Dec 17 2013

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.5% Tween/H2O for 15 min RT
Specification
Brain
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 16 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (brain)
Antigen retrieval step
None
Specification
brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5 · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Mrs. Francoise Geffroy

Verified customer

Submitted Apr 07 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate Gel)
Loading amount
15 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Mar 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cynomolgus monkey Tissue sections (Cerebrum & Cerebellum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization
No
Specification
Cerebrum & Cerebellum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Kathleen .

Verified customer

Submitted Dec 20 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (Olfactory stem cells)
Permeabilization
Yes - 0,1% Triton X100
Specification
Olfactory stem cells
Blocking step
BSA 3% + Goat serum 5% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 18 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Olfactory stem cells GFP)
Permeabilization
Yes - 0,1% Triton X100
Specification
Olfactory stem cells GFP
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 25 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (Olfactory stem cells)
Permeabilization
Yes - 0,1% Triton X100
Specification
Olfactory stem cells
Blocking step
BSA 3% + Goat serum 5% as blocking agent for 1 hour as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (Olfactory stem cells (neuronal indicution))
Permeabilization
Yes - 0,1% Triton X100
Specification
Olfactory stem cells (neuronal indicution)
Blocking step
BSA 3% + Goat serum 5% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Antoine Veron

Verified customer

Submitted Sep 20 2016

1-10 of 44 Abreviews or Q&A

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