Overview

  • Product name
    Anti-Bak antibody [Y164] (HRP)
    See all Bak primary antibodies
  • Description
    Rabbit monoclonal [Y164] to Bak (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Bak aa 1-100. The exact sequence is proprietary.

  • Positive control
    • WB: HeLa whole cell lysate. IHC-P: FFPE human colon (normal) tissue sections.
  • General notes

    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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Proclin
    Constituents: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity
    Affinity purified
  • Clonality
    Monoclonal
  • Clone number
    Y164
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab199516 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
IHC-P 1/125 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 25 kDa (predicted molecular weight: 23 kDa).

Target

  • Function
    In the presence of an appropriate stimulus, accelerates programmed cell death by binding to, and antagonizing the anti-apoptotic action of BCL2 or its adenovirus homolog E1B 19k protein. Low micromolar levels of zinc ions inhibit the promotion of apoptosis.
  • Tissue specificity
    Expressed in a wide variety of tissues, with highest levels in the heart and skeletal muscle.
  • Sequence similarities
    Belongs to the Bcl-2 family.
  • Domain
    Intact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.
  • Cellular localization
    Mitochondrion membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apoptosis regulator BAK antibody
    • BAK antibody
    • BAK like antibody
    • Bak NT antibody
    • BAK_HUMAN antibody
    • Bak1 antibody
    • Bcl 2 homologous antagonist/killer antibody
    • Bcl 2 like 7 protein antibody
    • Bcl-2 homologous antagonist/killer antibody
    • Bcl-2-like protein 7 antibody
    • BCL2 antagonist/killer 1 antibody
    • Bcl2 like 7 Protein antibody
    • Bcl2-L-7 antibody
    • BCL2L7 antibody
    • CDN1 antibody
    • Cell death inhibitor 1 antibody
    • MGC117255 antibody
    • MGC3887 antibody
    • NBak antibody
    • Pro apoptotic protein BAK antibody
    see all

Images

  • All lanes : Anti-Bak antibody [Y164] (HRP) (ab199516) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : BAK1 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 25 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    ab199516 was shown to specifically react with Bak in wild-type HAP1 cells as signal was lost in BAK1 knockout cells. Wild-type and BAK1 knockout samples were subjected to SDS-PAGE. Ab199516 was incubated overnight at 4°C at 1/5000 dilution. Blots were developed with ECL technique.

  • IHC image of Bak staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab199516 at 1/125 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative 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.

     

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Anti-Bak antibody [Y164] (HRP) (ab199516) at 1/5000 dilution + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 23 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab199516 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab199516 has not yet been referenced specifically in any publications.

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

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