Technical Data
Calpain I, Domain II (Mu Calpain, Protein Kinase C Activating Factor)
Calpain is a calcium-dependent thiol proteinase that is functionally active as a heterodimer
composed of a regulatory subunit and one of at least two large catalytic subunits (calpain 1 or calpain 2). In vitro, Calpain 1 ( -calpain) requires micromolar levels of calcium, while calpain 2 (M-calpain) requires millimolar levels of calcium for activation. The regulation of calpain in vivo is the subject of many current studies, which suggest that proteolytic activity is regulated posttranscriptionally by mechanisms such as calcium requirements, subcellular localization of the heterodimer, phosphorylation via the EGFR-Erk signaling cascade, endogenous inhibitors (calpastatin) and autoproteolytic cleavage (1). Calpastatin negatively regulates autoproteolytic cleavage of calpain between Gly27 and Leu28 (2). Calpain influences cell migration by modifying rather than degrading its substrates responsible for cell adhesion and cytoskeletal arrangement. Control of calpain activity has caught the attention of drug development
since limiting its activity could mute invasiveness of tumors or chronic inflammation (1).

Suitable for use in Immunocytochemistry, Western Blotting. Other applications not tested.

Recommended Dilution:
Immunocytochemistry: 1:25
Western Blot: 1:1000
Note: Not recommended for Immunoprecipitation.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

100ul-20CBlue IceBovineMouse
Purified bovine -calpain.
Supplied as a liquid, 0.05% sodium azide.
Recognizes an epitope in domain II (aa 245-265) of -calpain from several mammalian sources (e.g. human, rat, pig, bovine). Detects a band of ~80kDa by Western blot. Does not cross-react with m-calpain, n-calpain, calmodulin or calpastatin.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
General References:
1. Fox, J.E. (1999) Thromb. Haemost. 82(2): 385-391 (Review article). 2. Chan, S.L. et al., (1999) J. Neurosci. Res. 58(1):167-190 (Review article). 3. Huang. Y. and K.K. Wang (2001) Trends Mol.
Med. 7(8): 355-362 (Review article). 4. Glading, A., et al., (2002) Trends Cell Biol. 12(1): 46-54 (Review article). 5. Tidball, J.G. et al., (2000) Int. J. Biochem. Cell Biol. 32(1): 1-5 (Review article).