Revision 1

#3769

Store at -20C

BNIP3 Antibody
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W

Reactivity:
M R

Sensitivity:
Endogenous

MW (kDa):
22-28, 50-55

Source/Isotype:
Rabbit

UniProt ID:
#O55003

Entrez-Gene Id:
12176

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

BNIP3 Antibody detects endogenous levels of total BNIP3 protein from mouse and rat.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser60 of BNIP3 from mouse/rat. Antibodies were purified by peptide affinity chromatography.

Background

BNIP3 (Bcl-2/E1B-19kDa interacting protein 3) is a pro-apoptotic mitochondrial protein and Bcl-2 family member that contains a Bcl-2 homology 3 (BH3) domain and a carboxyl-terminal transmembrane (TM) domain (1-3). While BNIP3 has a predicted molecular weight of about 22 kDa, it runs anomalously on SDS-PAGE and includes a band of around 60 kDa that may be a dimeric form that is not reduced (2). BNIP3 associates with anti-apoptotic family members Bcl-2, Bcl-xL, and the adenovirus homologue E1B-19kDa. BNIP3 is distinct from other Bcl-2 family members that contain only the BH3 domain in that the TM domain, and not the BH3 domain, is required for mitochondrial targeting and pro-apoptotic activity (4). In addition to apoptosis, BNIP3 has been implicated in necrosis (5) and autophagy (6-11). In hypoxic conditions, BNIP3 can induce mitochondrial autophagy (mitophagy) by disrupting the Bcl-2-Beclin-1 complex (9). BNIP3 can also promote mitophagy by triggering the translocation of the E3 ubiquitin ligase Parkin to the mitochondria (10) or by directly binding LC3 on the autophagosome (11). BNIP3 may also localize to the endoplasmic reticulum (ER) where it can selectively induce the autophagic clearance of ER (ERphagy) (11). Increased expression of BNIP3 under hypoxic conditions is mainly regulated by the transcription factor HIF-1α (12-14). Silencing of the BNIP3 promoter by methylation has been observed in several types of cancer cells and may play an important role in their survival (14-18).

Background References

  1. Boyd, J.M. et al. (1994) Cell 79, 341-51.
  2. Chen, G. et al. (1997) J Exp Med 186, 1975-83.
  3. Yasuda, M. et al. (1998) J Biol Chem 273, 12415-21.
  4. Ray, R. et al. (2000) J Biol Chem 275, 1439-48.
  5. Vande Velde, C. et al. (2000) Mol Cell Biol 20, 5454-68.
  6. Daido, S. et al. (2004) Cancer Res 64, 4286-93.
  7. Tracy, K. et al. (2007) Mol Cell Biol 27, 6229-42.
  8. Quinsay, M.N. et al. (2010) Autophagy 6, 855-62.
  9. Bellot, G. et al. (2009) Mol Cell Biol 29, 2570-81.
  10. Lee, Y. et al. (2011) Am J Physiol Heart Circ Physiol 301, H1924-31.
  11. Hanna, R.A. et al. (2012) J Biol Chem 287, 19094-104.
  12. Bruick, R.K. (2000) Proc Natl Acad Sci USA 97, 9082-7.
  13. Guo, K. et al. (2001) Cell Death Differ 8, 367-76.
  14. Sowter, H.M. et al. (2001) Cancer Res 61, 6669-73.
  15. de Angelis, P.M. et al. (2004) Int J Oncol 24, 1279-88.
  16. Okami, J. et al. (2004) Cancer Res 64, 5338-46.
  17. Murai, M. et al. (2005) Clin Cancer Res 11, 1021-7.
  18. Murai, M. et al. (2005) Br J Cancer 92, 1165-72.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting

Cross-Reactivity Key

M: Mouse R: Rat

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

#3769

BNIP3 Antibody

Cell Signaling Technology Logo
Western blot analysis of extracts from C2C12 cells, untreated (-) or cobalt chloride-treated (100 μM; overnight) using BNIP3 Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: BNIP3 Antibody
Western blot analysis of HeLa cells, mock transfected or transfected with mouse BNIP3, CTLL-2, RAW 264.7 and BaF3 cells using BNIP3 Antibody.
Western Blotting Image 2: BNIP3 Antibody
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.