Recombinant P. humanus PINK1 Protein, CF Summary
Reaction conditions will need to be optimized for each specific application. We recommend an initial PINK1 concentration of 0.5-2 µM for the phosphorylation of recombinant Parkin, Ubiquitin, or Polyubiquitin chains.
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CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||X mg/ml (X μM) in 20 mM HEPES pH 7.6, 50 mM NaCl, 10% Glycerol (v/v), 1 mM TCEP|
|Shipping||The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
Serine/Threonine kinase PINK1 (PTEN-induced putative kinase protein 1) plays a critical role in preventing mitochondrial dysfunction during cellular stress. PINK is translated in the cytosol, then translocated to the outer mitochondrial membrane where it is rapidly cleaved and degraded as a part of normal mitochondrial function. In damaged (depolarized) mitochondria PINK becomes stabilized and accumulates, resulting in the subsequent phosphorylation of numerous proteins on the mitochondrial surface including Mfn2. Ultimately PARK2 (E3 Ubiquitin Ligase Parkin) is recruited to the damaged mitochondria where it is activated by 1) PINK-mediated phosphorylation of PARK2 at serine 65, and 2) PARK2 interaction with phosphorylated Ubiquitin (also phosphorylated by PINK on serine 65). This signaling cascade is critical for clearing the damaged mitochondria via selective autophagy (mitophagy) by mediating activation and translocation of PARK2.
Recombinant human PINK1 is not active in vitro, while this protein from the Human Body Louse (Pediculus humanus) effectively phosphorylates recombinant Parkin, mono-Ubiquitin, and poly-Ubiquitin chains. It specifically phosphorylates both Parkin and Ubiquitin at serine 65. This recombinant protein contains an N-terminal GST tag.
- Kane L.A., et al. (2014) J. Cell Biol. 205: 143
- Matsuda N., et al. (2010) J. Cell Biol. 189: 211
- Vives-Bauza C., et al. (2010) Proc. Natl. Acad. Sci. 107: 378
- Wauer T., et al. (2015) EMBO J. 34: 307
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