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Purified Mouse Anti- Calcineurin
Purified Mouse Anti- Calcineurin

Western blot analysis of calcineurin. Lysate from A-431 cells was probed with anti-calcineurin (clone G182-1847) at concentrations of 1.0 (lane 1), 0.2 (lane 2), and 0.04 µg/ml (lane 3). The 61 kDa A subunit of calcineurin is identified.

Western blot analysis of calcineurin. Lysate from A-431 cells was probed with anti-calcineurin (clone G182-1847) at concentrations of 1.0 (lane 1), 0.2 (lane 2), and 0.04 µg/ml (lane 3). The 61 kDa A subunit of calcineurin is identified.

Product Details
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BD Pharmingen™
Human (QC Testing)
Mouse IgG1
Human β1 isoform of calcineurin A Subunit aa. 349-505
Western blot (Routinely Tested), Immunoprecipitation (Tested During Development)
61 kDa
0.5 mg/ml
AB_396379
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

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.

Product Notices

  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.
556350 Rev. 8
Antibody Details
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G182-1847

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.

556350 Rev. 8
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
556350 Rev.8
Citations & References
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Development References (3)

  1. Fruman DA, Klee CB, Bierer BE, Burakoff SJ. Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proc Natl Acad Sci U S A. 1992; 89(9):3686-3690. (Biology). View Reference
  2. Husi H, Luyten MA, Zurini MG. Mapping of the immunophilin-immunosuppressant site of interaction on calcineurin. J Biol Chem. 1994; 269(19):14199-14204. (Biology). View Reference
  3. Lai MM, Burnett PE, Wolosker H, Blackshaw S, Snyder SH. Cain, a novel physiologic protein inhibitor of calcineurin. J Biol Chem. 1998; 273(29):18325-18331. (Biology). View Reference
556350 Rev. 8

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.