APPLICATION
Application Note
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 associated 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 100 μl were then transferred to TTN-coated microtiter wells and incubated for 2h at 37ºC. 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 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of CAPN3 and TTN was in a dose dependent manner.
Observed MW
32 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Ile602~Ala821 (NP_000061.1)
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
calpain 3 , CANP3 , CANPL3 , LGMD2 , LGMD2A , LGMDD4 , LGMDR1 , nCL-1 , p94
Background
Calpain, a heterodimer consisting of a large and a small subunit, is a major intracellular protease, although its function has not been well established. This gene encodes a muscle-specific member of the calpain large subunit family that specifically binds to titin. Mutations in this gene are associated 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. [provided by RefSeq, Jul 2008]
Database
Research Area
DATA IMAGES
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GTX00194-pro Functional Assay Image
Functional ELISA analysis of GTX00194-pro Human Calpain 3 protein which can bind immobilized TTN protein.
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GTX00194-pro Image
SDS-PAGE analysis of GTX00194-pro Human Calpain 3 protein.
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GTX00194-pro Image
WB analysis of GTX00194-pro Human Calpain 3 protein.
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REFERENCE
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REVIEW
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