Recombinant
RabMAb

Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

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Overview

  • Product name
    Anti-Alpha B Crystallin antibody [EPR2752]
    See all Alpha B Crystallin primary antibodies
  • Description
    Rabbit monoclonal [EPR2752] to Alpha B Crystallin
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Alpha B Crystallin (C terminal).

  • Positive control
    • Fetal heart and rat eyeball lysates; human heart muscle tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    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.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

Applications

Our Abpromise guarantee covers the use of ab76467 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 - 1/2000. Predicted molecular weight: 20 kDa.
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol). 
ICC/IF 1/250.
IP 1/250.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function
      May contribute to the transparency and refractive index of the lens.
    • Tissue specificity
      Lens as well as other tissues.
    • Involvement in disease
      Defects in CRYAB are the cause of myofibrillar alpha-B crystallin-related (MFM-CRYAB) [MIM:608810]. A neuromuscular disorder that results in weakness of the proximal and distal limb muscles, weakness of the neck, velopharynx and trunk muscles, hypetrophic cardiomyopathy, and cataract in a subset of patients.
    • Sequence similarities
      Belongs to the small heat shock protein (HSP20) family.
    • Cellular localization
      Cytoplasm. Nucleus. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles.
    • Information by UniProt
    • Database links
      • Alternative names
        • AACRYA antibody
        • Alpha B crystallin antibody
        • Alpha crystallin B chain antibody
        • Alpha(B)-crystallin antibody
        • Alpha-crystallin B chain antibody
        • CRYA2 antibody
        • Cryab antibody
        • CRYAB_HUMAN antibody
        • Crystallin alpha B antibody
        • Crystallin alpha polypeptide 2 antibody
        • CTPP2 antibody
        • Heat shock 20 kD like protein antibody
        • Heat shock protein beta 5 antibody
        • Heat shock protein beta-5 antibody
        • HspB5 antibody
        • Renal carcinoma antigen NY REN 27 antibody
        • Renal carcinoma antigen NY-REN-27 antibody
        • Rosenthal fiber component antibody
        see all

      Anti-Alpha B Crystallin antibody [EPR2752] images

      • Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling Alpha B Crystallin with purified ab76467 at 1/250. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        Immunohistochemical staining of paraffin embedded human brain with purified ab76467 at a working dilution of 1 in 250. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

      • Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Anti-Alpha B Crystallin antibody [EPR2752] (ab76467) at 1/1000 dilution (purified) + Rat eyeball lysate at 10 µg

        Secondary
        HRP goat anti-rabbit (H+L) at 1/1000 dilution

        Predicted band size : 20 kDa
        Observed band size : 22 kDa (why is the actual band size different from the predicted?)

        Blocking buffer: 5% NFDM/TBST

        Dilution buffer: 5% NFDM/TBST

      • Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Anti-Alpha B Crystallin antibody [EPR2752] (ab76467) at 1/1000 dilution (purified) + Human fetal heart tissue lysate at 10 µg/ml

        Secondary
        HRP goat anti-rabbit (H+L) at 1/1000 dilution

        Predicted band size : 20 kDa
        Observed band size : 22 kDa (why is the actual band size different from the predicted?)

        Blocking buffer: 5% NFDM/TBST

        Dilution buffer: 5% NFDM/TBST

      • Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        Immunofluorescence staining of HeLa cells with purified ab76467 at a working dilution of 1 in 200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 200. The cells were fixed in 4% PFA.

      • Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)


        Predicted band size : 20 kDa

        ab76467 (purified) at 1/250 immunoprecipitating alpha B crystallin in human fetal heart (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

        Blocking buffer and concentration: 5% NFDM/TBST.

        Diluting buffer and concentration: 5% NFDM /TBST.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        Immunohistochemical staining of paraffin embedded human brain with unpurified ab76467 at a working dilution of 1 in 250. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        Immunohistochemical staining of alpha B Crystallin in paraffin embedded human heart muscle with ab76467 at a 1/100 dilution.

      • Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Western blot - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Lane 1 : Anti-Alpha B Crystallin antibody [EPR2752] (ab76467) at 1/5000 dilution
        Lane 2 : Anti-Alpha B Crystallin antibody [EPR2752] (ab76467) at 1/20000 dilution

        Lane 1 : fetal heart lysate
        Lane 2 : rat eyeball lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        goat anti-rabbit HRP at 1/2000 dilution

        Predicted band size : 20 kDa
        Observed band size : 20 kDa
      • Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunocytochemistry/ Immunofluorescence - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
      • Immunoprecipitation - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)
        Immunoprecipitation - Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

        ab76467 (unpurified) at 1/250 immunoprecipitating alpha B crystallin in human fetal heart (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

        Blocking buffer and concentration: 5% NFDM/TBST.

        Diluting buffer and concentration: 5% NFDM /TBST.

      References for Anti-Alpha B Crystallin antibody [EPR2752] (ab76467)

      ab76467 has not yet been referenced specifically in any publications.

      Publishing research using ab76467? Please let us know so that we can cite the reference in this datasheet.

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      Submit an abreview for ab76467 Submit a question for ab76467

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      Western blot abreview for Anti-alpha B Crystallin antibody [EPR2752]

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      Abreviews
      Abcam guarantees this product to work in the species/application used in this Abreview.
      Application
      Western blot
      Sample
      Rat Cell lysate - whole cell (cardiomyocytes)
      Loading amount
      8 µg
      Specification
      cardiomyocytes
      Gel Running Conditions
      Reduced Denaturing (10% gel)
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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      Submitted Feb 10 2010

      Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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