Anti-Calcineurin A (CnA)

Calcineurin is a Ca/calmodulin-dependent serine-threonine phosphatase that plays an important role in transducing Ca-dependent signals in a variety of cell types. Calcineurin has also been shown to have a profound influence on the properties of striated muscle cells, including cardiac muscle. Calcineurin (also known as CALNA or CALNA1, Calcineurin-alpha, Protein phosphatase 2B or PP2B) is the Ca+/calmodulin-regulated protein phosphatase, first detected in skeletal muscle and brain, has been found in from yeast to mammals. It is a heterodimers of two subunits: Calcineurin B/CnB, the 19-kda Ca+-binding and regulatory subunit, and Calcineurin A/CnA, ~61-kda catalytic subunit that is highly homologous with PP1 and PP2A. Multiple catalytic subunits of calcineurin are derived from at least 2 structural genes, type 1 (calcineurin A-alpha) and type 2 (calcineurin A-beta, CALNA2), each of which can produce additional alternatively spliced transcripts. CnB belongs to the family of EF-hand Ca-binding proteins. Both CnB and calmodulin are important for the activation of the phosphatase activity of calcineurin. Calcineurin controls the production of many cytokines including IL-2, TNF-alpha in the T-cell activation pathway. Calcineurin mediated dephosphorylation of the nuclear factor of activated T-cells (NF-AT) is required for NF-AT activation, nuclear translocation, and subsequent gene expression in T-cells. The immunosuppressive drugs, such as FK506, inhibit activation of NF-AT by calcineurin. Human calcineurin A-alpha (~60kD) and calcineurin B (~19kD) were co-expressed with yeast myristoyl-CoA:protein N-myrisoyltransferase and purified to >95%. Recombinant and purified Calcineurin is N-myristoylated on the CnB, as in the natural protein, resulting into biologically active protein. For western blot +ve control , calcineurin is supplied in SDS-PAGE sample buffer (reduced). Load ~10 ul/lane to visualize with antibodies. , Applications: , Suitable for use in ELISA. Western Blot, though not tested, may potentially be used as an application. Other applications not tested. Recommended Dilution: Western Blot: 1-10ug/ml using ECL. B subunit is ~19kD., ELISA: 1:10,000-1:100,000 using 50-100ng of control peptide/well., Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. 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.