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RecombinantRabMAb

Recombinant Anti-GFAP antibody [EPR19996] (ab207165)

Reviews (2) Submit a question References (6)
Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
  • Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
  • Immunoprecipitation - Anti-GFAP antibody [EPR19996] (ab207165)
  • Anti-GFAP antibody [EPR19996] (ab207165)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19996] to GFAP
  • Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB, IHC-Fr, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-GFAP antibody [EPR19996]
    See all GFAP primary antibodies

  • Description

    Rabbit monoclonal [EPR19996] to GFAP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB, IHC-Fr, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Flow Cyt: Mouse primary brain cell. ICC/IF: Rat hippocampal mixed glia.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207165 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
Flow Cyt (Intra)
1/500.
ICC/IF (1)
Use a concentration of 0.1 - 1 µg/ml.
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/5000. Detects a band of approximately 40-54 kDa (predicted molecular weight: 49 kDa).
IHC-Fr (1)
1/100.

Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20)
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.
ICC/IF
Use a concentration of 0.1 - 1 µg/ml.
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/5000. Detects a band of approximately 40-54 kDa (predicted molecular weight: 49 kDa).
IHC-Fr
1/100.

Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20)
IP
1/30.

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

    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

  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR19996] (ab207165)
    Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR19996] (ab207165)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary brain cells cells labelling GFAP with ab207165 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 [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of human cerebrum [PMID: 15378652] is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labelling GFAP with ab207165 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in mouse primary astrocyte. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (4 µg/mL) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
  • Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunofluorescence staining of GFAP using ab207165 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 ab207165 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 ab207165 gave comparable results using 4% formaldehyde fixation (10 min).

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab207165 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic and membrane staining on glial cells of mouse cerebrum is observed. 

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
    Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
    Lane 1 : Anti-GFAP antibody [EPR19996] (ab207165) at 1/10000 dilution
    Lane 2 : Anti-GFAP antibody [EPR19996] (ab207165) at 1/1000 dilution

    Lane 1 : Human brain lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 40-54 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the literature.

    Negative control: NIH/3T3 (PMID: 824020; PMID:2294; PMID: 6340792; PMID: 9466565).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on tumor cells of human glioma is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of mouse cerebrum is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Specific cytoplasmic staining on myenteric nerve plexus, and negative on epithelial cells and smooth muscle cells of mouse colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of rat hippocampus is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Specific cytoplasmic staining on myenteric nerve plexus, and negative on epithelial cells and smooth muscle cells of rat colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
    Western blot - Anti-GFAP antibody [EPR19996] (ab207165)
    All lanes : Anti-GFAP antibody [EPR19996] (ab207165) at 1/5000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate
    Lane 3 : Rat cerebellum lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 49 kDa
    Observed band size: 40-54 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the literature (PMID: 824020; PMID:2294; PMID: 6340792).

  • Immunoprecipitation - Anti-GFAP antibody [EPR19996] (ab207165)
    Immunoprecipitation - Anti-GFAP antibody [EPR19996] (ab207165)

    GFAP was immunoprecipitated from 0.35 mg of human brain lysate with ab207165 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207165 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Human brain lysate 10µg (Input).

    Lane 2: ab207165 IP in human brain lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207165 in human brain lysate.

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

    Exposure time: 1 second.

  • Anti-GFAP antibody [EPR19996] (ab207165)
    Anti-GFAP antibody [EPR19996] (ab207165)

References (6)

Publishing research using ab207165? Please let us know so that we can cite the reference in this datasheet.

ab207165 has been referenced in 6 publications.

  • Tribble JR  et al. Retinal Ganglion Cell Degeneration in a Rat Magnetic Bead Model of Ocular Hypertensive Glaucoma. Transl Vis Sci Technol 10:21 (2021). PubMed: 33510960
  • Rossino MG  et al. A Nature-Inspired Nrf2 Activator Protects Retinal Explants from Oxidative Stress and Neurodegeneration. Antioxidants (Basel) 10:N/A (2021). PubMed: 34439544
  • Cammalleri M  et al. Novel Insights into Beta 2 Adrenergic Receptor Function in the rd10 Model of Retinitis Pigmentosa. Cells 9:N/A (2020). PubMed: 32917020
  • Ji S  et al. Increased hippocampal TrkA expression ameliorates cranial radiation-induced neurogenesis impairment and cognitive deficit via PI3K/AKT signaling. Oncol Rep 44:2527-2536 (2020). PubMed: 33125501
  • Kudo T  et al. Autoimmune Glial Fibrillary Acidic Protein Astrocytopathy Presenting with Slowly Progressive Myelitis and Longitudinally Extensive Spinal Cord Lesions. Intern Med 59:2777-2781 (2020). PubMed: 32669494
  • Cammalleri M  et al. Efficacy of a Fatty Acids Dietary Supplement in a Polyethylene Glycol-Induced Mouse Model of Retinal Degeneration. Nutrients 9:N/A (2017). PubMed: 28961167

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFAP antibody [EPR19996]

 Excellent
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.

Abcam user community

Verified customer

Submitted Apr 21 2021

Immunohistochemistry (Frozen sections) abreview for Anti-GFAP antibody [EPR19996]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
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.

Abcam user community

Verified customer

Submitted Oct 21 2019

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