Description
Calcineurin is a Ca2+- and calmodulin-dependent serine/threonine phosphatase which consists of two subunits. The A subunit (~61 kDa) is catalytic and has calmodulinbinding activity; the B subunit (~19 kDa), which is myristylated at its N-terminus, has Ca[2+]-binding activity. Calcineurin is expressed ubiquitously in eukaryotic cells, including yeast. The protein is highly expressed in mammalian brain, and has been implicated in a number of calcium-dependent pathways in the nervous system, including regulation of neurotransmitter release. Calcineurin also plays a key role in T cell receptor-mediated signal transduction pathways. Calcineurin dephosphorylates components of NF-AT (Nuclear Factor of Activated T cells), allowing NF-AT to translocate to the nucleus where it participates with AP-1 in activation of multiple cytokine and surface receptor genes. Inhibition of calcineurin by immunosuppressive agents such as cyclosporin A and FK506 correlates with the suppression of early T cell activation events. These immunosuppressive agents bind to specific intracellular proteins, called immunophilins, to form complexes that inhibit calcineurin's phosphatase activity. In the presence of these complexes, NF-AT remains phosphorylated and thus cannot translocate to the nucleus. A novel protein, Cain (calcineurin inhibitor), has been described which displays a similar pattern of neuronal and brain expression and has been shown to bind and inhibit calcineurin activity in vivo. G182-1847 recognizes the α and β isoforms of the A subunit of calcineurin (~61 kDa). It reacts with an epitope in the autoinhibitory domain. It does not recognize the B subunit of calcineurin. A recombinant tag-fusion protein containing amino acids 349-505 of the human β1 isoform of calcineurin A subunit was used as immunogen.
Applications include immunoprecipitation (1-2 µg/ml) and western blot analysis (1-2 µg/ml). Jurkat human T cells (ATCC TIB-152)2, HeLa human carcinoma cells (ATCC CCL-2), and A-431 human epidermal carcinoma cells (ATCC CRL-1555) are suggested as positive controls. A431 cell lysate (Cat. No. 611447) and HeLa cell lysate (Cat. No. 611449) are also available as ready-to-use positive western blot control.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
