Revision 1

#44060Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

22-28, 50-55

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q12983

Entrez-Gene Id:

664

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:100
Immunofluorescence (Immunocytochemistry) 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

BNIP3 (D7U1T) Rabbit mAb recognizes endogenous levels of total BNIP3 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human BNIP3 protein.

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).

  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

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#44060

BNIP3 (D7U1T) Rabbit mAb

Western Blotting Image 1: BNIP3 (D7U1T) Rabbit mAb Expand Image
Western blot analysis of extracts from HeLa or MCF7 cells, untreated (-) or cobalt chloride-treated (100 μM, overnight; +), using BNIP3 (D7U1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: BNIP3 (D7U1T) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human BNIP3 protein (hBNIP3-Myc; +), using BNIP3 (D7U1T) Rabbit mAb.
Immunoprecipitation Image 1: BNIP3 (D7U1T) Rabbit mAb Expand Image
Immunoprecipitation of BNIP3 from MCF7 cells treated with cobalt chloride (100 μM; overnight). Lane 1 represents 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BNIP3 (D7U1T) Rabbit mAb. Western blot analysis was performed using BNIP3 (D7U1T) Rabbit mAb. A conformation-specific secondary antibody was used to avoid reactivity with IgG.
Immunohistochemistry Image 1: BNIP3 (D7U1T) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using BNIP3 (D7U1T) Rabbit mAb.
Immunohistochemistry Image 2: BNIP3 (D7U1T) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using BNIP3 (D7U1T) Rabbit mAb.
Immunohistochemistry Image 3: BNIP3 (D7U1T) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded MCF7 cell pellets, untreated (left) or cobalt chloride-treated (right), using BNIP3 (D7U1T) Rabbit mAb.
Immunohistochemistry Image 4: BNIP3 (D7U1T) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal human liver using BNIP3 (D7U1T) Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).
Immunofluorescence Image 1: BNIP3 (D7U1T) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with cobalt chloride (100 μM, 24 hr; right), using BNIP3 (D7U1T) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalliodin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).