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

    Synthetic peptide within Human Bid aa 50-150. The exact sequence is proprietary.

  • 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

  • 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

    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

  • 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:
  • Zhang B  et al. Metastasis-associated protein 2 promotes the metastasis of non-small cell lung carcinoma by regulating the ERK/AKT and VEGF signaling pathways. Mol Med Rep 17:4899-4908 (2018). Read more (PubMed: 29393472) »
  • Yan W  et al. Fisetin inhibits the proliferation of gastric cancer cells and induces apoptosis through suppression of ERK 1/2 activation. Oncol Lett 15:8442-8446 (2018). Read more (PubMed: 29805580) »
See all 13 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your reply. In terms of rabbit antibodies to Bid that have been tested in ICC, we have only ab32060 https://www.abcam.com/Bid-antibody-Y8-ab32060.html It has not been tested in mouse yet, though. The immunogen is a synthetic peptide corresponding to residues before the BH3 domain of human Bid (it is amino acids 50-62). So this will not recognize the p15 fragment resulting from cleavage at 61/62 or p11 or p13. See http://www.uniprot.org/uniprot/P55957 We also have a mouse monoclonal that was tested in ICC/IF (ab114051), but not in mouse. It was raised against a purified recombinant fragment of Human Bid expressed in E. coli, but I have no further information right now about the sequence covered by the immunogen. It seems that we might not have the exact product you are looking for. I might suggest checking also www.biocompare.com. I just wanted to point out that we do have a testing discount program for UNTESTED species and/or applications. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. 

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

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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