Overview

  • Product nameAnti-Bak antibody [Y164]
    See all Bak primary antibodies
  • Description
    Rabbit monoclonal [Y164] to Bak
  • Specificityab32371 recognises Bak. The antibody does not cross-react with other Bcl2 members.
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive)

  • Positive control
    • Hela cell lysate, human stomach carcinoma
  • General notes

     

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityMonoclonal
  • Clone numberY164
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32371 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 - 1/5000. Predicted molecular weight: 23 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/10 - 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/250 - 1/500.
IP 1/100.

Target

  • FunctionIn 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 specificityExpressed in a wide variety of tissues, with highest levels in the heart and skeletal muscle.
  • Sequence similaritiesBelongs to the Bcl-2 family.
  • DomainIntact 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 localizationMitochondrion 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

Anti-Bak antibody [Y164] images

  • ICC/IF image of ab32371 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab32371, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) 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.

  • ab32371 staining Bak in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Anti-Bak antibody [Y164] (ab32371) at 1/5000 dilution + HeLa cell lysate

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
  • Immunohistochemical analysis of Bak expression in paraffin embedded human stomac carcinoma, using 1/250 ab32371.


  • Predicted band size : 23 kDa

    See Abreview

References for Anti-Bak antibody [Y164] (ab32371)

This product has been referenced in:
  • Holloway A  et al. Resistance to UV-induced apoptosis by ß-HPV5 E6 involves targeting of activated BAK for proteolysis by recruitment of the HERC1 ubiquitin ligase. Int J Cancer 136:2831-43 (2015). Read more (PubMed: 25408501) »
  • Wang B  et al. Role of Ku70 in deubiquitination of Mcl-1 and suppression of apoptosis. Cell Death Differ 21:1160-9 (2014). Read more (PubMed: 24769731) »

See all 16 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (COLO205 colon carcinoma xenograft)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification COLO205 colon carcinoma xenograft
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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Submitted Dec 14 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample Chinese Hamster Cell (CHO cell)
Specification CHO cell
Permeabilization Yes - 1% TRITON-X-100
Fixative Paraformaldehyde
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Submitted Mar 18 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (MEF)
Specification MEF
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Jul 30 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample Rat Cell (h9c2)
Specification h9c2
Permeabilization Yes - 1% TRITON-X-100
Fixative Paraformaldehyde
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Submitted Jul 30 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample Mouse Cell (MEF)
Specification MEF
Permeabilization Yes - 1% TRITON-X-100
Fixative Paraformaldehyde
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Submitted Jul 30 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (h9c2)
Specification h9c2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Jul 30 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Immuno-precipitation step Protein A/G
Sample Human Cell lysate - whole cell (HCT116 p53-/- cell line)
Specification HCT116 p53-/- cell line
Total protein in input 100 µg
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Mr. Christian Marx

Verified customer

Submitted Aug 27 2013

The concentration of 1542-1 lot YI063010CS is 0.1160 mg/ml.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (isolated mitochondria from NB4 cell line)
Loading amount 30 µg
Specification isolated mitochondria from NB4 cell line
Gel Running Conditions Reduced Denaturing (13%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. Christian Marx

Verified customer

Submitted Feb 15 2013

Thank you for contacting us.  The purpose of the blocking step is to prevent non-specific binding between the antibodies and the tissue section, membrane, 96-well plate, etc.  It is possible to get a good signal without a blocking step, but blocking wi...

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