- References (2)
- Inventor Info
|Vector Type||CAPN1/pET and CAPNS1deltaGR/pACpET|
|Antigen/Gene or Protein Targets||Calpain-1, µ-Calpain, Calpain 1, Calpain1|
Calpain belongs to the superfamily of Ca2+-regulated cysteine proteases, which are indispensable to the regulation of various cellular functions. Of the 15 mammalian calpain isoforms, µ- and m-calpains are the best characterized. Both µ- and m-calpain are ubiquitously expressed and exist as heterodimers, containing a distinct 80-kDa catalytic subunit (CAPN1 and CAPN2, respectively) and the common, 30-kDa regulatory subunit (CAPNS1).
Calpain-1 (µ-Calpain) Plasmid is an E.coli expression system for recombinant human µ-calpain. As detailed in Hata et al., 2013 this is achieved by co-expressing CAPN1 with a N-terminal His tag and CAPNS1 lacking the N-terminal Gly-rich (GR) domain (CAPNS1deltaGR) in the SoluBL21 E.coli strain. From 1 L of E. coli culture, over 2 and 6 mg, respectively, of µ-calpain and its active-site mutant µ-calpain:C115S (CAPN1:C115S+CAPNS1deltaGR) were purified by two successive column chromatographies. When the enzyme is compared to the native enzyme, the purified µ-calpain shows almost identical properties, demonstrating its suitability for use in structural and functional studies.
|Research Area||Cell Signaling & Signal Transduction|
There are 2 reference entries for this reagent.View All References
Dr Hiroyuki Sorimachi
from Tokyo Metropolitan Institute Of Medical Science