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Recombinant

Recombinant Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

Submit a review Submit a question References (3)
Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
  • Western blot - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
  • Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [EPR1034Y] to GFAP - Mouse IgG2a
  • Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra), ICC
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric)
    See all GFAP primary antibodies

  • Description

    Mouse monoclonal [EPR1034Y] to GFAP - Mouse IgG2a

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra), ICCmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • IHC-P: Human cerebral cortex tissue. WB: Human, mouse and rat brain tissue lysate. IHC-Fr: Mouse cerebrum tissue. Flow Cyt (intra): Rat primary neural/glia cells. IP: Rat brain tissue lysate. ICC: mouse neural/glia cells

  • General notes

    This mouse monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab68428). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab279290 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
1/1000.
IHC-Fr
1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
IP
1/30.
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.
ICC
1/50.
Notes
WB
1/1000.
IHC-Fr
1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
IP
1/30.
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.
ICC
1/50.

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

  • Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

    Immunocytochemical analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized mouse neural/glia tissue labeling GFAP with ab279290 at 1/50 (16 µg/ml) dilution followed by ab98736 Goat Anti-Mouse IgG (DyLight® 488) pre-adsorbed at 1/1000 dilution (Green). Positive staining on mouse primary astrocytes is observed. The nuclear counterstain was DAPI (Blue).

    Counterstain: ab207165 anti-GFAP rabbit mAb at 1/1000 dilution with secondary antibody ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution.

    Negative controls: ab279290 with secondary ab150080 at 1/500 dilution; and ab207165 at 1/1000 dilution with secondary ab98376 at 1/1000 dilution

     

  • Western blot - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Western blot - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    All lanes : Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290) at 1/1000 dilution

    Lane 1 : Human brain tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Observed band size: 40-50 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

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

    Exposure times: Lane 1: 3.25 seconds; Lane 2: 48 seconds; Lane 3: 10 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

    IHC image of GFAP staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BONDTM 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 20mins. The section was then incubated with ab279290, 1µg/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG2a, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit 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.

  • Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab279290 at 1/500 (1.634 µg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

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

  • Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neural/glia cells labelling GFAP with ab279290 at 1/1000 dilution (0.1µg)/ Right compared with a Mouse monoclonal IgG isotype control/ Left. Goat Anti-Mouse IgG (Alexa Fluor® 647, ab150119) at 1/2000 dilution was used as the secondary antibody. 

  • Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)
    Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

    GFAP was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 µg with ab279290 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279290 at 1/1000 dilution. mouse IgG for IP (HRP) (ab131368) was used at 1/5000 dilution.

    Lane 1: Rat brain tissue lysate  10µg.

    Lane 2: ab279290 IP in rat brain tissue lysate.

    Lane 3: Mouse monoclonal IgG2a (ab18413) instead of ab279290 in rat brain tissue lysate.

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

    Exposure time: 23 seconds.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (3)

Publishing research using ab279290? Please let us know so that we can cite the reference in this datasheet.

ab279290 has been referenced in 3 publications.

  • Li J  et al. Sertad1 Induces Neurological Injury after Ischemic Stroke via the CDK4/p-Rb Pathway. Mol Cells 45:216-230 (2022). PubMed: 35014620
  • Dong P  et al. Roles of ERRα and TGF-β signaling in stemness enhancement induced by 1 µM bisphenol A exposure via human neural stem cells. Exp Ther Med 23:164 (2022). PubMed: 35069845
  • Sengupta S  et al. Differentiated glioma cell-derived fibromodulin activates integrin-dependent Notch signaling in endothelial cells to promote tumor angiogenesis and growth. Elife 11:N/A (2022). PubMed: 35642785

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