Revision 2

#43110Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
SQSTM1/p62 (D5E2) Rabbit mAb 8025 20 µl 62 kDa Rabbit IgG
NDP52 (D1E4A) Rabbit mAb 60732 20 µl 52, 60 kDa Rabbit IgG
Optineurin (D2L8S) Rabbit mAb 58981 20 µl 75 kDa Rabbit IgG
Parkin (Prk8) Mouse mAb 4211 20 µl 50 kDa Mouse IgG2b
PINK1 (D8G3) Rabbit mAb 6946 20 µl 60, 50 kDa Rabbit IgG
BNIP3 (D7U1T) Rabbit mAb 44060 20 µl 22-28, 50-55 kDa Rabbit IgG
BNIP3L/Nix (D4R4B) Rabbit mAb 12396 20 µl 38, 76 kDa Rabbit IgG
LC3B (D11) XP® Rabbit mAb 3868 20 µl 14, 16 kDa Rabbit IgG
Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb 62802 20 µl Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Mitophagy Antibody Sampler Kit provides an economical means of detecting proteins involved in the process of mitophagy. The kit includes enough primary antibody to perform two western blot experiments with each primary antibody.

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 antibodies.

Background

Autophagy is a catabolic process for the autophagosome-lysosomal degradation of bulk cytoplasmic contents (1, 2). Selective autophagy targets the degradation of distinct sets of substrates and organelles (3-5). One of the best studied examples of selective autophagy involves the clearance of damaged mitochondria through a process called mitophagy. Several pathways have been described for various contexts of mitophagy, including the FUNDC1 pathway, the BNIP3 and BNIP3L/Nix pathway, and the PINK1/Parkin pathway. FUNDC1 is a mitochondrial protein that is phosphorylated by the autophagy kinase ULK1 and regulates hypoxia induced mitophagy (6, 7). BNIP3L/Nix and BNIP3 are members of the Bcl-2 family of apoptosis regulators that are expressed on mitochondria, induced by hypoxia, and have been shown to play a role in mitophagy (8). BNIP3L/Nix is also important in the autophagic maturation of erythroid cells (9). FUNDC1, BNIP3 and BNIP3L/Nix bind to LC3 family members, targeting the mitochondria to the autophagosome.

Non-hypoxic induction of mitophagy can be regulated by the PINK1/Parkin pathway, which plays causative roles in neurodegenerative disease, most notably Parkinson’s disease (10, 11). PINK1 is a mitochondrial serine/threonine kinase that is stabilized on the outer mitochondrial membrane of damaged mitochondria. Substrates of PINK1 include the E3 ubiquitin ligase Parkin and ubiquitin itself (12-14). Phosphorylation of Parkin as well as binding to phosphorylated ubiquitin leads to accumulation of ubiquitinated chains on multiple mitochondrial proteins. Ubiquitinated proteins are recognized by selective cargo receptors including SQSTM1/p62, Optineurin, and NDP52 (15-16). Autophagy cargo receptors contain an LC3-interacting region (LIR) required for binding to Atg8/LC3 family members and targeting to the autophagosome (3).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  3. Birgisdottir, Å.B. et al. (2013) J Cell Sci 126, 3237-47.
  4. Xu, Z. et al. (2015) Acta Biochim Biophys Sin (Shanghai) 47, 571-80.
  5. Mancias, J.D. and Kimmelman, A.C. (2016) J Mol Biol 428, 1659-80.
  6. Liu, L. et al. (2012) Nat Cell Biol 14, 177-85.
  7. Wu, W. et al. (2014) EMBO Rep 15, 566-75.
  8. Sowter, H.M. et al. (2001) Cancer Res 61, 6669-73.
  9. Sandoval, H. et al. (2008) Nature 454, 232-5.
  10. Kitada, T. et al. (1998) Nature 392, 605-8.
  11. Valente, E.M. et al. (2004) Science 304, 1158-60.
  12. Kim, Y. et al. (2008) Biochem Biophys Res Commun 377, 975-80.
  13. Kane, L.A. et al. (2014) J Cell Biol 205, 143-53.
  14. Koyano, F. et al. (2014) Nature 510, 162-6.
  15. Heo, J.M. et al. (2015) Mol Cell 60, 7-20.
  16. Lazarou, M. et al. (2015) Nature 524, 309-314.

Background References

    Trademarks and Patents

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