Revision 3

#12396Store 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, IF-IC

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

38, 76

Source/Isotype:

Rabbit IgG

UniProt ID:

#O60238

Entrez-Gene Id:

665

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400

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

BNIP3L/Nix (D4R4B) Rabbit mAb recognizes endogenous levels of total BNIP3L/Nix protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu128 of human BNIP3L/Nix protein.

Background

BCL2/Adenovirus E1B 19 kDa protein-interacting protein 3-like (BNIP3L) (1), also termed BNIP3α (2), B5 (3), and Nix (4), is a member of the Bcl-2 family of apoptotic regulators with highest homology to BNIP3. BNIP3L can bind BNIP3, Bcl-xL, and Bcl-2 (1-5). BNIP3L forms homodimers that withstand denaturing by SDS and reducing conditions (5). BNIP3L is a mitochondrial protein and knockout studies suggest that BNIP3L regulates autophagic clearance of damaged mitochondria during erythroid maturation via mitochondrial autophagy (6,7). It has been shown that the expression of BNIP3L is up-regulated during terminal erythroid differentiation (6-8), as well as in tumor cell lines during hypoxia (9-11). BNIP3L directly regulates the elimination of mitochondria through its ability to bind to and recruit important components of the autophagic machinery, including LC3/Atg8 and GABARAP proteins, via its amino-terminal LC3-interacting region (LIR) (12). BNIP3L may also indirectly activate phagophore formation either via the recruitment of autophagy proteins or by binding Bcl-xL, which in turn releases Beclin-1 (13). BNIP3L/Nix also plays a pivotal role in Parkin-mediated mitochondrial autophagy via its ability to mediate the mitochondrial translocation of Parkin (14). Activated BNIP3L can promote the opening of mitochondrial permeability transition pores resulting in mitochondrial depolarization, generation of reactive oxygen species, and induction of necrosis. Due to its involvement in cell death and autophagy, research scientists have implicated BNIP3L in heart disease and cancer (13).

  1. Matsushima, M. et al. (1998) Genes Chromosomes Cancer 21, 230-5.
  2. Yasuda, M. et al. (1999) Cancer Res 59, 533-7.
  3. Ohi, N. et al. (1999) Cell Death Differ 6, 314-25.
  4. Chen, G. et al. (1999) J Biol Chem 274, 7-10.
  5. Imazu, T. et al. (1999) Oncogene 18, 4523-9.
  6. Schweers, R.L. et al. (2007) Proc Natl Acad Sci USA 104, 19500-5.
  7. Sandoval, H. et al. (2008) Nature 454, 232-5.
  8. Aerbajinai, W. et al. (2003) Blood 102, 712-7.
  9. Sowter, H.M. et al. (2001) Cancer Res 61, 6669-73.
  10. Bruick, R.K. (2000) Proc Natl Acad Sci USA 97, 9082-7.
  11. Fei, P. et al. (2004) Cancer Cell 6, 597-609.
  12. Novak, I. et al. (2010) EMBO Rep 11, 45-51.
  13. Zhang, J. and Ney, P.A. (2009) Cell Death Differ 16, 939-46.
  14. Ding, W.X. et al. (2010) J Biol Chem 285, 27879-90.

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 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.
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 3
#12396

BNIP3L/Nix (D4R4B) Rabbit mAb

Western Blotting Image 1: BNIP3L/Nix (D4R4B) Rabbit mAb Expand Image
Western blot analysis of A172 and HeLa cells, untreated (-) or cobalt chloride-treated (100 μM, overnight; +), using BNIP3L/Nix (D4R4B) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: BNIP3L/Nix (D4R4B) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using BNIP3L/Nix (D4R4B) Rabbit mAb.
Western Blotting Image 3: BNIP3L/Nix (D4R4B) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing DYKDDDDK-tagged full-length human BNIP3L/Nix (hBNIP3L/Nix-DYKDDDDK; +), using BNIP3L/Nix (D4R4B) Rabbit mAb. The BNIP3L/Nix construct was kindly provided by Dr. Reuben Shaw, Salk Institute for Biological Studies, La Jolla, CA.
Immunoprecipitation Image 1: BNIP3L/Nix (D4R4B) Rabbit mAb Expand Image
Immunoprecipitation of BNIP3L/Nix from A172 cells, treated with cobalt chloride (100 μM, overnight), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or BNIP3L/Nix (D4R4B) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using BNIP3L/Nix (D4R4B) Rabbit mAb.
Immunofluorescence Image 1: BNIP3L/Nix (D4R4B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of A172 cells, untreated (left) or cobalt chloride-treated (100 μM, overnight; right), using BNIP3L/Nix (D4R4B) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).