Overview

  • Product name
  • Description
    Rabbit polyclonal to Bak
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Bak.

  • Positive control
    • This antibody gave a positive signal in A431 Whole Cell Lysate.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92999 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use at an assay dependent concentration. 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

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Bak knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab92999 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab92999 was shown to recognize Bak in wild-type HAP1 cells as signal was lost at the expected MW in Bak knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Bak knockout samples were subjected to SDS-PAGE. ab92999 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-Bak antibody (ab92999) at 1 µg/ml + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    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?)
    Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • ICC/IF image of ab92999 stained HepG2 cells. The cells were 4% formaldehyde fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab92999 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit IgG (H+L)(ab96899) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

ab92999 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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