Overview

  • Product nameAnti-Bid antibody [Y8]
    See all Bid primary antibodies
  • Description
    Rabbit monoclonal [Y8] to Bid
  • SpecificityThis antibody does not cross-react with other Bcl-2 family members.
  • Tested applicationsSuitable 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

    This product is a recombinant rabbit monoclonal antibody.

    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.

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

    Alternative versions available:

    Anti-Bid antibody (HRP) [Y8] (ab201754)

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

  • FunctionThe 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 specificityIsoform 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).
  • 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.
  • 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 localizationCytoplasm; 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

Anti-Bid antibody [Y8] images

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

    Predicted band size : 22 kDa
    Observed band size : 22 kDa
  • Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using ab32060 at 1/100 dilution.
  • All lanes : Anti-Bid antibody [Y8] (ab32060) at 1/500 dilution

    Lane 1 : Whole cell lysate prepared from breast cancer clinical extract
    Lane 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
    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

    Image courtesy of an anonymous Abreview.

    See Abreview

  • 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 for Anti-Bid antibody [Y8] (ab32060)

This product has been referenced in:
  • 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) »
  • Tao LJ  et al. Tetrandrine induces apoptosis and triggers a caspase cascade in U2-OS and MG-63 cells through the intrinsic and extrinsic pathways. Mol Med Rep 9:345-9 (2014). WB ; Human . Read more (PubMed: 24173687) »

See all 7 Publications for this product

Product Wall

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
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
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Submitted Jul 01 2011

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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Submitted Feb 21 2011

Application Flow Cytometry
Sample Human Cell (U937)
Specification U937
Fixation Leucoperm Reagent A
Permeabilization Yes - Leucoperm Reagent B
Gating Strategy Live Cells
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Submitted Sep 09 2008

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
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Submitted Sep 03 2008

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