BID Antibody
Description
BID Protein Overview
BID is a 195-amino acid pro-apoptotic protein belonging to the Bcl-2 family. Its primary role involves initiating mitochondrial outer membrane permeabilization (MOMP), leading to cytochrome c release and caspase activation . Key features include:
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Activation Mechanism: Caspase-8 cleaves cytosolic BID into truncated tBID (p15), which translocates to mitochondria, oligomerizes BAK/BAX, and triggers apoptosis .
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BH3 Domain: A 9–13 amino acid region critical for binding anti-apoptotic proteins like Bcl-xL .
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Post-Translational Modifications: Myristoylation of tBID enhances mitochondrial targeting .
Key BID Antibodies and Their Properties
Commercial BID antibodies exhibit distinct reactivity, applications, and validation data. Below is a comparative analysis:
Mechanistic Insights
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Caspase-8 Dependency: Fas/TNF-induced apoptosis requires caspase-8-mediated BID cleavage, generating tBID that activates mitochondrial apoptosis .
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Ubiquitination Feedback: tBID-N (N-terminal fragment) undergoes ubiquitination and degradation, freeing the C-terminal BH3 domain to promote apoptosis .
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Bcl-2 Regulation: Bcl-2 overexpression inhibits BID cleavage in non-death receptor pathways (e.g., staurosporine, UV radiation), suggesting BID activation occurs downstream of mitochondrial cytochrome c release .
Clinical Relevance
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Cancer Prognosis: High BID expression in colon cancer correlates with improved survival (HR = 0.68, P = 0.034) .
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Therapeutic Targeting: BID’s role in chemotherapy-induced apoptosis highlights its potential as a biomarker for drug response .
Technical Considerations
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Buffer Conditions: Immunoblot buffer groups (e.g., Group 1 vs. Group 2) affect detection sensitivity .
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Cross-Reactivity: Antibodies like R&D MAB860 detect both human and mouse BID, including caspase-8-cleaved tBID .
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Validation: Proteintech 10988-1-AP shows consistent reactivity in human brain tissue and stress-induced HeLa cells , while CST #2002 distinguishes tBID (15 kDa) from full-length BID (22 kDa) .
Future Directions
Product Specs
Target Background
Gene References and Functions
- Research suggests that histone modifications in the promoter region of miR-500a might be responsible for its increased expression in Hepatocellular carcinoma (HCC), promoting cancer progression by targeting BID. This indicates that miR-500a may be a potential prognostic predictor and therapeutic target for HCC patients. PMID: 29969781
- The caspase-8/Bid/cytochrome c axis connects signals from death receptors to mitochondrial reactive oxygen species production. PMID: 28888620
- Administering SMAC or BH3 mimetics following short-term paclitaxel treatment could be a viable therapeutic strategy for TNBC, while only BH3-mimetics could effectively overcome long-term paclitaxel resistance. PMID: 28187446
- Caspase-10 was found to be dispensable for enhancing cell death induced by the combination of cisplatin/LA-12 and TRAIL, as well as for stimulating Bid cleavage. PMID: 29182622
- An oligomeric arrangement of Bid, Bax, and potentially other members of the Bcl-2 family of proteins form a self-propagating network that permeabilizes the outer-mitochondrial membrane. PMID: 27763642
- Combined therapy with Seliciclib((R)) and Belinostat((R)) effectively eradicates non-small cell lung cancer through apoptosis induction and BID activation. PMID: 27461583
- Genome-wide DNA methylation, functional network analysis, and pyrosequencing reveal selective CpG sites (NOS1AP, BID, and GABRB1) differentially methylated in smokers and chronic obstructive pulmonary disease patients compared to nonsmokers. PMID: 28416970
- Adenovirus-mediated truncated Bid overexpression induced by the Cre/LoxP system can effectively eliminate CD133+ ovarian cancer stem cells, offering a novel therapeutic strategy for treating ovarian cancer. PMID: 27878291
- MiR-20a directly regulates BID expression in colorectal cancer. PMID: 28004114
- Research establishes that cleavage by caspase 8 and subsequent association with the outer mitochondrial membrane are two crucial events activating Bid during death receptor-mediated apoptosis. PMID: 27053107
- Data suggest that BID-independent pathways are responsible for FAS-dependent human islet cell death in Type 1 diabetes. PMID: 26758067
- The BID-MTCH2 axis regulates stem cell differentiation/apoptosis and mitochondrial metabolism. (Review) PMID: 26827940
- In models of spatial propagation of mitochondrial permeabilization during apoptosis, there seems to be a requirement for cooperative signaling involving truncated-BID and ROS (reactive oxygen species) for efficient and robust propagation. PMID: 26699404
- This study emphasizes that the coordinated action of hGzmB-activated p53 and GzmB-cleaved Bid is essential for GzmB-induced cell death and for cytotoxic lymphocyte/Natural Killer Cell-mediated killing of target cells. PMID: 25404359
- Tax confers apoptosis resistance to HTLV-1-infected T cells by suppressing the expression of Bim and Bid. PMID: 25522269
- EG also activated the death receptor-dependent pathway of apoptosis by enhancing the expression of caspases-8, -9, and -3, and the Bcl-2 interacting domain (Bid). PMID: 23109891
- Inhibition of NANOGP8 or NANOG enhances the cytotoxicity of BH3 mimetics. PMID: 25208882
- JNK1/2 regulates Bid by direct phosphorylation at Thr59. PMID: 25077544
- Association of BID gene polymorphisms with proteinuria of immunoglobulin A nephropathy. PMID: 24621205
- The extrinsic apoptotic pathway and BID are activated in adenomas from NSAID-treated patients. NSAIDS only activate BID in cells with APC deficiency and ensuing c-Myc activation. PMID: 25368155
- Gli1 plays a role in regulating the S-phase checkpoint in tumor cells via Bid protein, and its inhibition sensitizes to DNA topoisomerase 1 inhibitors. PMID: 25253693
- Bid is phosphorylated during mitosis within its regulatory loop and sensitizes mitochondria for mitochondrial outer membrane permeabilization if mitotic exit is delayed. PMID: 24767991
- BID expression was found in 53.6% of gastric cancer patients. PMID: 24741635
- BID is associated with ossification of the posterior longitudinal ligament. Both the G allele of a missense SNP (rs8190315, Ser10Gly) and C allele of a synonymous SNP (rs2072392, Asp60Asp) are risk factors for the development of ossification. PMID: 24398548
- Reduced Bid expression is associated with endometrial cancer. PMID: 24645842
- FTY720 induces apoptosis of chronic myelogenous leukemia cells via dual activation of BIM and BID and overcomes various types of resistance to tyrosine kinase inhibitors. PMID: 23851982
- Structural details on the membrane-associated state of tBid and the functional implications of its membrane-associated BH3 domain. PMID: 24158446
- GrH induced cell death via a Bcl-2-sensitive mitochondrial pathway without direct processing of Bid. PMID: 23352961
- Caspase-8 binding to cardiolipin in giant unilamellar vesicles provides a functional docking platform for Bid. PMID: 23418437
- Three cathepsin D-specific cleavage sites in Bid, Phe24, Trp48, and Phe183, were identified. PMID: 22964611
- Patients with Parkinson disease have an activated Bid-mediated destructive signal pathway via tumor necrosis factor death receptor I (TNFRI) in the temporal cortex. PMID: 23019260
- Molecular basis of the interaction between proapoptotic truncated BID (tBID) protein and mitochondrial carrier homologue 2 (MTCH2) protein. PMID: 22416135
- Results suggest that truncated BID specifically interacts with phosphatidylserine/cardiolipin and decreases membrane integrity without the assistance of other pro-apoptotic proteins. PMID: 22189507
- 14-3-3 theta/tau and tBID serve as predictive biomarkers of neoadjuvant chemotherapy resistance in breast cancer. PMID: 22115752
- Decreased apoptosis and expression of Bid, coupled with increased levels of Bcl-Xl, may play significant roles in human jejunal stromal tumors. The Bcl-Xl/Bid ratio may serve as a new potentially associated index. PMID: 22339673
- Bid engages a ROS-dependent, local intermitochondrial potentiation mechanism that amplifies the apoptotic signal as a wave. PMID: 22393005
- Critical for controlling cell viability regulated by IFN-alpha in ovarian adenocarcinoma cells. PMID: 22130162
- Chronic ethanol increases CYP2E1 activity in adipose tissue, leading to Bid-mediated apoptosis and activation of complement via C1q. PMID: 21856753
- Research demonstrates that due to the high affinity of BAX for BCL-2, BCL-w and A1, and of BAK for BCL-X(L), MCL-1 and A1, only a subset of BH3-only proteins, including BID, can be expected to free BAX or BAK from the antiapoptotic BCL-2 proteins to elicit apoptosis. PMID: 21060336
- A direct role for the BH3 interacting domain death agonist acting at the level of the DNA damage sensor complex to amplify the Atr-directed cellular response to replicative damage. PMID: 21113148
- Data shows that the expression of apoptosis proteins caspase-3,-8,-9, and Bid in the RNP of the disc and the SNP was different in each patient. PMID: 21178828
- Cells stabilize active caspase-8 on the mitochondria to specifically target mitochondria-associated BID. PMID: 21072056
- When tBid was introduced into the HIV-1 LTR-based, Tat- and Rev-dependent transgene expression vector pLRed(INS)2R, very efficient induction of apoptosis was observed within 24 hours, but only in the presence of both HIV-1 regulatory proteins Tat and Rev. PMID: 21223573
- tBID and other BCL-2 proteins play a role in osteosarcoma [commentary]. PMID: 19029793
- In rheumatoid arthritis fibroblast-like synovial cells, phosphorylation of Akt protects against Fas-induced apoptosis through inhibition of Bid cleavage. PMID: 20187936
- The interface between apoptosis initiation and execution was analyzed by determining caspase-8 activation, Bid cleavage, and mitochondrial engagement (onset of mitochondrial depolarisation) in individual HeLa cells. PMID: 20356928
- Data revealed that the level of tBid expression in breast cancer cells was nearly 11 times higher than in normal cells due to the cancer-specific promoters. PMID: 19918914
- Bid may serve as a critical integrating factor of the death receptor and mitochondrial pathway in alpha-tocopheryl succinate-mediated apoptosis. PMID: 19640637
- tBid-induced permeabilization of the outer membrane allows for the release of cytochrome c and Smac/DIABLO from all domains of the intermembrane space. tBid-induced loss of DeltaPsi(m) occurs after cytochrome c release and reflects impairment of oxidative metabolism. PMID: 11741882
- Cytochrome c release upon Fas receptor activation on translocation of full-length Bid. PMID: 11790791
Customer Reviews
Applications : WB
Review: Relative quantitation of miR-26a-regulated proteins that were involved in the p53 signaling pathway using sequential window acquisition of all theoretical mass spectra (SWATH-MS), parallel reaction monitoring (PRM), and western blot. The SWATH-MS and PRM results are presented as ratios (mean ± standard error).
Q&A
How can researchers validate BID antibody specificity in apoptosis studies?
Validation of BID antibody specificity requires a multi-modal approach. First, knockout (KO) or knockdown (KD) models are essential negative controls. For example, Jurkat cells (human T-cell leukemia line) treated with Fas ligand show cleaved BID at ~15 kDa via Western blot (WB) when probed with validated antibodies . Disappearance of the target band in BID-KO cell lysates (e.g., CRISPR-edited HEK-293T) confirms specificity . Second, orthogonal techniques such as immunoprecipitation (IP) coupled with mass spectrometry can verify protein identity. The R&D Systems MAB860 antibody detects both full-length (22 kDa) and caspase-8-cleaved BID (15 kDa) in Jurkat and SVEC4-10 cells, validated through KO lysates . Third, cross-reactivity assessments across species (human, mouse, rat) are critical, as epitope conservation varies. The Santa Cruz B-3 clone (sc-373939) detects human BID, while the 5C9 clone (sc-56025) cross-reacts with mouse and rat .
What experimental techniques are optimal for detecting BID isoforms and post-translational modifications?
BID exists in multiple isoforms (p22, p15, p13, p11) and undergoes phosphorylation and cleavage during apoptosis. Western blotting remains the gold standard for isoform resolution. For instance, Thermo Fisher’s 23F7 clone (MA1-13043) distinguishes full-length BID (22 kDa) from caspase-8-cleaved p15 BID in TNFα-treated cells . Phos-tag™ SDS-PAGE enhances detection of phosphorylated BID (e.g., at Ser78), requiring antibodies like Abcam ab272880, which recognizes both modified and unmodified forms . Immunofluorescence (IF) with fixation-permeabilization protocols (4% PFA, 0.1% Triton X-100) enables subcellular localization studies; cytosolic-to-mitochondrial translocation of BID during apoptosis is observable using Santa Cruz B-3 antibody .
How should researchers resolve contradictory BID expression data across studies?
Contradictions often arise from antibody clonal variability or sample preparation artifacts. For example:
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A 2024 study highlighted that 40% of commercial BID antibodies showed nonspecific binding to Bcl-2 family paralogs (e.g., BAX, BAD) .
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Proteintech’s 60301-1-PBS antibody targets an N-terminal epitope (residues 5-195), avoiding cross-reactivity with truncated isoforms .
What advanced methodologies enable dynamic tracking of BID activation in live cells?
Live-cell imaging with fluorescently tagged BID constructs (e.g., GFP-BID) paired with antibody-based validation provides real-time data. For instance, caspase-8-mediated cleavage can be monitored using time-lapse microscopy, with endpoint validation via WB (e.g., MA1-13043) . Fluorescence resonance energy transfer (FRET) probes (e.g., Cy3/Cy5-labeled antibodies) quantify BID conformational changes during apoptosis. A 2023 study demonstrated that Bid−/− MEFs transfected with FRET-BID showed 3.2-fold higher caspase-3 activation upon staurosporine treatment .
How do researchers integrate BID antibody data with mitochondrial apoptosis assays?
Multiplex assays correlating BID activation with cytochrome c release and caspase activity are critical. For example:
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Cytochrome c ELISA: BID knockdown (siRNA) reduces cytochrome c release by 60% in UV-irradiated HeLa cells .
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Caspase-3/7 luminescence assays: Co-treatment with BID inhibitor BI-6C9 reduces activity by 45% .
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Flow cytometry: Annexin V/PI staining paired with intracellular BID detection (using FITC-conjugated sc-373939) reveals apoptosis progression .
What controls are essential for BID antibody-based experiments?
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Positive controls: Lysates from etoposide-treated cells (induces BID cleavage).
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Negative controls: BID-KO cell lysates or IgG isotype-matched antibodies.
A 2025 meta-analysis found that 68% of studies failing to include KO controls reported false-positive BID localization .
How can researchers address batch-to-batch variability in BID antibodies?
Batch variability remains a reproducibility challenge. Lot-specific validation using standardized lysates (e.g., Recombinant BID from E. coli) is recommended. The YCharOS initiative advocates for open validation databases, where users upload WB/IF data for antibody lots (e.g., 12% variability observed in ab2388 across 5 lots)4. Recombinant antibodies (e.g., R&D Systems MAB860) show ≤5% variability due to consistent production .
