Bcl-2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3), also known as 19 kDa interacting protein 3 (NIP3), is a proapoptotic member of Bcl-2 protein family. BNIP3 is a 194 amino acid, 21.5 kDa (predicted) protein that contains a single Bcl-2 homology 3 (BH3) domain and a C-terminal transmembrane domain required for mitochondrial localization, homodimerization, and regulation of its proapoptotic function. BNIP3 was identified as one of several proteins that interact with discrete domains of Bcl-2 and the E1B 19 kDa protein. Under conditions of prolonged oxygen deprivation, the hypoxia-induced protein HIF1-alpha activates expression of BNIP3, which in turn, promotes apoptosis under these conditions. The mechanism of BNIP3-mediated apoptosis is independent of caspase activation and cytochrome c release and is characterized by early plasma membrane and mitochondrial damage, prior to the appearance of chromatin condensation or DNA fragmentation. Human BNIP3 shares 90% amino acid identity with mouse and rat BNIP3. Human BNIP3 shares 56% amino acid sequence identity with human BNIP3L.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Western Blot
Cited:
Immunohistochemistry, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human BNIP3
Ser2-Glu160
Accession # Q12983
Ser2-Glu160
Accession # Q12983
Specificity
Detects human BNIP3 in Western blots. In Western blots, this antibody shows no cross-reactivity with recombinant human BNIP3L.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human BNIP3 Antibody
Detection of Human BNIP3 by Western Blot.
Western blot shows lysates of A549 human lung carcinoma cell line and MCF-7 human breast cancer cell line untreated (-) or treated (+) with 150 µM CoCl2for 16 hours. PVDF membrane was probed with 1 µg/mL of Human BNIP3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4147), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BNIP3 at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.Detection of BNIP3 by Western Blot
The hypoxic pathway in glioblastoma tumours. Levels of 7 HIF-pathway members were estimated by Western blotting (A), with densitometry measures (B), or measured by ELISA (C). A HIF-pathway score was derived for each patient by combining the relative scores of 7 hypoxia-responsive proteins (D); IDH1 mutant samples are shown as solid square symbols. n = 37 samples; T, tumour, +, positive control (MDA-MB-231 cell line exposed to 1% O2 for 16 h), mw, molecular weight marker; mean ± SEM. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35419292), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human BNIP3 Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: CoCl2-treated A549 human lung carcinoma cell line and MCF‑7 human breast cancer cell line
Sample: CoCl2-treated A549 human lung carcinoma cell line and MCF‑7 human breast cancer cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: BNIP3
Long Name
BCL2/Adenovirus E1B 19 Kda Protein-Interacting Protein 3-Like
Alternate Names
Nip3
Gene Symbol
BNIP3
UniProt
Additional BNIP3 Products
Product Documents for Human BNIP3 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human BNIP3 Antibody
For research use only
Related Research Areas
Citations for Human BNIP3 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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