Overview

  • Product name

    Anti-Bak antibody [AT38E2]
    See all Bak primary antibodies
  • Description

    Mouse monoclonal [AT38E2] to Bak
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to amino acids 29-187 of Human Bak, purified from E. coli.

  • Positive control

    • WB: Mouse lung lysate, 293T, HeLa, A431, A549, Jurkat, MCF7, and PC3 cell lysates ICC/IF: HeLa cells. FC: HeLa cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 24/01/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    AT38E2
  • Isotype

    IgG2a
  • Light chain type

    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab104124 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
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 23 kDa.
ICC/IF Use at an assay dependent concentration.

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 - ab104124 observed at 0 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab104124 was found to be non-specific when BAK knockout samples were used. Wild-type and BAK knockout samples were subjected to SDS-PAGE. Ab104124 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/250 dilution 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.

    This image was produced using the ascites version of this antibody. 

  • Flow cytometry analysis of BAK in HeLa cells. The cell was stained with ab104124 at 2-5µg for 1x10cells (red). A Goat anti mouse IgG (Alexa fluor 488) was used as the secondary antibody. Mouse monoclonal IgG was used as the isotype control (blue), cells without incubation with primary and secondary antibody was used as the negative control (black).

  • Immunocytochemistry/ Immunofluorescence analysis of BAK in HeLa cells. The cell was stained with ab104124 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
  • All lanes : Anti-Bak antibody [AT38E2] (ab104124) at 1/1000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : A549 cell lysate
    Lane 5 : Jurkat cell lysate
    Lane 6 : MCF7 cell lysate
    Lane 7 : PC3 cell lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : goat anti-mouse secondary antibody conjugated to HRP

    Predicted band size: 23 kDa

  • Anti-Bak antibody [AT38E2] (ab104124) at 1/500 dilution + Mouse lung lyasate at 35 µg

    Secondary
    Goat anti-mouse HRP conjugate

    Developed using the ECL technique.

    Predicted band size: 23 kDa



    This image was produced using the ascites version of this antibody. 

  • Immunofluorescence staining of Bak in HeLa cells using Hoechst 3342 (Blue) for nucleus staining and ab104124 antibody (1/500) with Texas Red (Red).

    This image was produced using the ascites version of this antibody. 

References

This product has been referenced in:

  • Singh A  et al. Podophyllotoxin and Rutin Modulates Ionizing Radiation-Induced Oxidative Stress and Apoptotic Cell Death in Mice Bone Marrow and Spleen. Front Immunol 8:183 (2017). Read more (PubMed: 28289414) »
  • Liu P  et al. Downregulation of microRNA-125a is involved in intervertebral disc degeneration by targeting pro-apoptotic Bcl-2 antagonist killer 1. Iran J Basic Med Sci 20:1260-1267 (2017). Read more (PubMed: 29299204) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 16 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 16 2016

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