Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y8] to Bid
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Human
Product nameAnti-Bid antibody [Y8]
See all Bid primary antibodies
DescriptionRabbit 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 reactivityReacts with: Human
Synthetic peptide within Human Bid aa 50-150. The exact sequence is proprietary.
- Jurkat cell lysate and human prostate carcinoma.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
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.
|WB||1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
modificationsTNF-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
- Apoptic death agonist antibody
- Apoptotic death agonist BID antibody
- BH3 interacting domain death agonist antibody
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.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
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
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.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32060 has been referenced in 18 publications.
- Kabir S et al. The CUL5 ubiquitin ligase complex mediates resistance to CDK9 and MCL1 inhibitors in lung cancer cells. Elife 8:N/A (2019). PubMed: 31294695
- Bao L et al. MicroRNA-500a Promotes the Progression of Hepatocellular Carcinoma by Post-Transcriptionally Targeting BID. Cell Physiol Biochem 47:2046-2055 (2018). PubMed: 29969781
- 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). 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). PubMed: 29805580
- Yao TH et al. Propranolol induces hemangioma endothelial cell apoptosis via a p53-BAX mediated pathway. Mol Med Rep 18:684-694 (2018). PubMed: 29767244