Recombinant
RabMAb

Anti-Bid antibody [Y8] (ab32060)

Abreviews (4)Q&A (1)Specific References (10)

Overview

  • Product name
    Anti-Bid antibody [Y8]
    See all Bid primary antibodies
  • Description
    Rabbit monoclonal [Y8] to Bid
  • Host species
    Rabbit
  • Specificity
    This antibody does not cross-react with other Bcl-2 family members.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic peptide corresponding to residues before the BH3 domain of human Bid.

  • Positive control
    • Jurkat cell lysate and human prostate 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.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32060 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. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/10.

Target

  • Function
    The major proteolytic product p15 BID allows the release of cytochrome c (By similarity). Isoform 1, isoform 2 and isoform 4 induce ICE-like proteases and apoptosis. Isoform 3 does not induce apoptosis. Counters the protective effect of Bcl-2.
  • Tissue specificity
    Isoform 2 and isoform 3 are expressed in spleen, bone marrow, cerebral and cerebellar cortex. Isoform 2 is expressed in spleen, pancreas and placenta (at protein level). Isoform 3 is expressed in lung, pancreas and spleen (at protein level). Isoform 4 is expressed in lung and pancreas (at protein level).
  • 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.
  • Post-translational
    modifications
    TNF-alpha induces a caspase-mediated cleavage of p22 BID into a major p15 and minor p13 and p11 products.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
    p15 BID is ubiquitinated by ITCH; ubiquitination results in proteasome-dependent degradation.
  • Cellular localization
    Cytoplasm; Cytoplasm. Mitochondrion membrane. When uncleaved, it is predominantly cytoplasmic; Mitochondrion membrane. A significant proportion of isoform 2 localizes to mitochondria, it may be cleaved constitutively; Mitochondrion membrane. Associated with the mitochondrial membrane and Mitochondrion membrane. Translocates to mitochondria as an integral membrane protein.
  • Information by UniProt
  • Database links
    • Alternative names
      • Apoptic death agonist antibody
      • Apoptotic death agonist BID antibody
      • BH3 interacting domain death agonist antibody
      • BH3 interacting domain death agonist p11 antibody
      • BH3 interacting domain death agonist p13 antibody
      • BH3 interacting domain death agonist p15 antibody
      • BH3-interacting domain death agonist p11 antibody
      • BID antibody
      • BID isoform ES(1b) antibody
      • BID isoform L(2) antibody
      • BID isoform Si6 antibody
      • BID_HUMAN antibody
      • Desmocollin type 4 antibody
      • FP497 antibody
      • Human BID coding sequence antibody
      • MGC15319 antibody
      • MGC42355 antibody
      • p11 BID antibody
      • p13 BID antibody
      • p15 BID antibody
      • p22 BID antibody
      see all

    Images

    • Western blot - Anti-Bid antibody [Y8] (ab32060)
      Western blot - Anti-Bid antibody [Y8] (ab32060)
      Anti-Bid antibody [Y8] (ab32060) at 1/1000 dilution + Jurkat cell lysate

      Predicted band size: 22 kDa
      Observed band size: 22 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bid antibody [Y8] (ab32060)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bid antibody [Y8] (ab32060)
      Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ab32060 at 1/100 dilution.
    • Western blot - Anti-Bid antibody [Y8] (ab32060)
      Western blot - Anti-Bid antibody [Y8] (ab32060)Image courtesy of an anonymous Abreview.
      All lanes : Anti-Bid antibody [Y8] (ab32060) at 1/500 dilution

      Lanes 1-2 : Whole cell lysate prepared from breast cancer clinical extract
      Lane 3 : Whole cell lysate prepared from MCF7 cells

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG-HRP at 1/1000 dilution

      Developed using the ECL technique.

      Predicted band size: 22 kDa
      Observed band size: 22 kDa


      Exposure time: 25 minutes

      See Abreview

    • Flow Cytometry - Anti-Bid antibody [Y8] (ab32060)
      Flow Cytometry - Anti-Bid antibody [Y8] (ab32060)
      Overlay histogram showing HeLa cells stained with ab32060 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32060, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References

    This product has been referenced in:
    • Wang C  et al. Bifidobacterial recombinant thymidine kinase-ganciclovir gene therapy system induces FasL and TNFR2 mediated antitumor apoptosis in solid tumors. BMC Cancer 16:545 (2016). WB ; Human . Read more (PubMed: 27464624) »
    • Stefanowicz-Hajduk J  et al. Pennogenyl Saponins from Paris quadrifolia L. Induce Extrinsic and Intrinsic Pathway of Apoptosis in Human Cervical Cancer HeLa Cells. PLoS One 10:e0135993 (2015). WB ; Human . Read more (PubMed: 26295969) »

    See all 10 Publications for this product

    Publishing research using ab32060? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    5 reviews or Q&A

    10
    Votes: Highest First

    The preferred option would be to p15. But we need to be tested in immunocytochemistry.

    Thank you for your reply. In terms of rabbit antibodies to Bid that have been tested in ICC, we have only ab32060 http://www.abcam.com/Bid-antibody-Y8-ab32060.html It has not been tested in mouse yet, though. The immunogen is a synthetic peptide corres...

    Read More

    Western blot abreview for Anti-Bid antibody [Y8]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Breast Cancer)
    Loading amount
    20 µg
    Specification
    Breast Cancer
    Gel Running Conditions
    Reduced Denaturing (12 %)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
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    Abcam user community

    Verified customer

    Submitted Jul 01 2011

    Western blot abreview for Anti-Bid antibody [Y8]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Oral Cancer)
    Loading amount
    20 µg
    Specification
    Oral Cancer
    Gel Running Conditions
    Non-reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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    Verified customer

    Submitted Feb 21 2011

    Flow Cytometry abreview for Anti-Bid antibody [Y8]

     Poor
    Abreviews
    Application
    Flow Cytometry
    Sample
    Human Cell (U937)
    Specification
    U937
    Fixation
    Leucoperm Reagent A
    Permeabilization
    Yes - Leucoperm Reagent B
    Gating Strategy
    Live Cells
    Read More
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    Abcam user community

    Verified customer

    Submitted Sep 09 2008

    Western blot abreview for Anti-Bid antibody [Y8]

     Inconclusive
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U937 and PBMC)
    Loading amount
    4e+006 cells
    Specification
    U937 and PBMC
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    2% Milk with 1% BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
    Read More
    Username

    Abcam user community

    Verified customer

    Submitted Sep 03 2008

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

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