Active Recombinant Human CAPN3 Protein, His-tagged

• Cat.No.: CAPN3-22H
• Product Overview: Recombinant Human CAPN3 Protein is produced by E.coli expression system with a His tag at the N-terminal.

Specification

• Description: Calpain 3 is a calcium-dependent cysteine protease mainly expressed in skeletal muscle. In humans, calpain 3 is encoded by the CAPN3 gene. This gene encodes a muscle-specific member of the calpain large subunit family that specifically binds to titin. Mutations in this gene are asscentigradeiated with limb-girdle muscular dystrophies type 2A. Alternate promoters and alternative splicing result in multiple transcript variants encoding different isoforms and some variants are ubiquitously expressed. Besides, Titin (TTN) has been identified as an interactor of CAPN3, thus a binding ELISA assay was conducted to detect the interaction of recombinant human CAPN3 and recombinant human TTN. Briefly, CAPN3 were diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to TTN-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CAPN3 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50μL stop solution to the wells and read at 450nm immediately. The binding activity of CAPN3 and TTN was shown in Figure 1, and this effect was in a dose dependent manner.
• Source: E. coli
• Species: Human
• Tag: His
• Form: 20 mM Tris, 150 mM NaCl, pH 8.0, containing 0.05 % sarcosyl and 5 % trehalose.
• Bio-activity: Binding activity of CAPN3 with TTN
• Molecular Mass: 32 kDa
• Protein length: Ile602-Ala821
• AA Sequence: IIFVSDRANSNKELGVDQESEEGKG KTSPDKQKQSPQPQPGSSDQESEEQ QQFRNIFKQIAGDDMEICADELKKV LNTVVNKHKDLKTHGFTLESCRSMI ALMDTDGSGKLNLQEFHHLWNKIKA WQKIFKHYDTDQSGTINSYEMRNAV NDAGFHLNNQLYDIITMRYADKHMN IDFDSFICCFVRLEGMFRAFHAFDK DGDGIIKLNVLEWLQLTMYA
• Endotoxin: <1.0EU per 1μg (determined by the LAL method).
• Purity: >95%
• Applications: Cell culture; Activity Assays.
• Notes: The kit is designed for in vitro and research use only, we will not be responsible for any issue if the kit was used in clinical diagnostic or any other prcentigradeedures.
• Stability: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37centigrade for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.
• Storage: Avoid repeated freeze/thaw cycles.
  Store at 2-8 centigrade for one month.
  Aliquot and store at -80 centigrade for 12 months.
• Reconstitution: Reconstitute in 20 mM Tris, 150 mM NaCl (pH 8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

Gene Information

• Gene Name: CAPN3 calpain 3, (p94) [ Homo sapiens ]
• Official Symbol: CAPN3
• Synonyms: CAPN3; calpain 3, (p94); LGMD2, LGMD2A; calpain-3; CANP3; nCL 1; p94; calpain L3; new calpain 1; LGMD2; nCL-1; CANPL3; LGMD2A; MGC4403; MGC10767; MGC11121; MGC14344
• Gene ID: 825
• mRNA Refseq: NM_000070
• Protein Refseq: NP_000061
• MIM: 114240
• UniProt ID: P20807