Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)

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 - Chimeric – BSA and Azide free
  • Suitable for: IHC-Fr, IP, IHC-P, WB, Flow Cyt (Intra), ICC
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free
    See all GFAP primary antibodies

  • Description

    Mouse monoclonal [EPR1034Y] to GFAP - Chimeric – BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-Fr, IP, IHC-P, WB, 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

    • 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. IHC-P: Human cerebral cortex tissue. ICC: Primary hippocampal rat neurons/glia

  • General notes

    ab279301 is the carrier free version of ab279289.

    This mouse 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.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Images

  • Western blot - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)
    Western blot - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)
    All lanes : Anti-GFAP antibody [EPR1034Y] - Chimeric (ab279289) 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


    This data was produced using ab279289, the same clone in a different formulation.

    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] - Chimeric – BSA and Azide free (ab279301)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)

    This data was produced using ab279289, the same clone in a different formulation.

    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 ab279289, 1µg/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG1, ab125913, 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] - Chimeric – BSA and Azide free (ab279301)
    Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)

    This data was produced using ab279289, the same clone in a different formulation.

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab279289 at /500 (1.968 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 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] - Chimeric – BSA and Azide free (ab279301)
    Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)

    This data was produced using ab279289, the same clone in a different formulation.

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized rat primary neural/glia cells labelling GFAP with ab279289 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] - Chimeric – BSA and Azide free (ab279301)
    Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] - Chimeric – BSA and Azide free (ab279301)

    This data was produced using ab279289, the same clone in a different formulation.

    GFAP was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 µg with ab279289 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279289 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: ab279289 IP in rat brain tissue lysate.

    Lane 3: Mouse monoclonal IgG1 (ab18443) instead of ab279289 in rat brain tissue lysate.

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

    Exposure time: 23 seconds.

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