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Vector Type CAPN1/pET and CAPNS1deltaGR/pACpET
Antigen/Gene or Protein Targets Calpain-1, µ-Calpain, Calpain 1, Calpain1
Relevance 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

References

There are 2 reference entries for this reagent.

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References: 2 entries

Hata et al. 2013. Genes Cells. 18(9):753-63. PMID: 23786391.

Efficient expression and purification of recombinant human μ-calpain using an Escherichia coli expression system.

Europe PMC ID: 23786391


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References: 2 entries

Hata et al. 2013. Genes Cells. 18(9):753-63. PMID: 23786391.

Efficient expression and purification of recombinant human μ-calpain using an Escherichia coli expression system.


Add a reference