Overview

  • Product name

    Cellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric)
    See all Calcineurin kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Enzyme activity
  • Product overview

    Calcineurin Activity Assay Kit (Colorimetric) ab139464 is designed for measuring cellular calcineurin (CaN, PP2B) phosphatase activity. It employs a convenient 96-well microtiter-plate format with all reagents necessary for measuring calcineurin (PP2B) phosphatase activity in tissue/cellular extracts.


    The detection of free-phosphate released in the calcineurin assay protocol is based on the classic Malachite green assay and offers the following advantages: non-radioactive; convenient 1-step detection; excellent sensitivity.


    This kit includes human recombinant calcineurin as a positive control. The RII phosphopeptide substrate (Calcineurin Substrate), supplied with this calcineurin assay, is the most efficient and outstanding peptide substrate known for calcineurin.

  • Notes

    Previously called Cellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric).

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    2X EGTA Buffer 1 x 1ml
    96-well Clear Microplate (1/2 Volume) 2 units
    Active Calcineurin Enzyme 1 x 10µl
    Calcineurin Assay Buffer 1 x 20ml
    Calcineurin Substrate 1 x 1.5mg
    Calmodulin 1 x 100µl
    Desalting Column 1 unit
    Desalting Resin 1 x 1g
    Green Assay Reagent 1 x 20ml
    Lysis Buffer 1 x 40ml
    Okadaic Acid 1 x 325µl
    Phosphate Standard 1 x 0.5ml
    Protease Inhibitor Cocktail 1 x 2 tablets
  • Research areas

  • Function

    Calcium-dependent, calmodulin-stimulated protein phosphatase. This subunit may have a role in the calmodulin activation of calcineurin. Dephosphorylates DNM1L, HSPB1 and SSH1.
  • Sequence similarities

    Belongs to the PPP phosphatase family. PP-2B subfamily.
  • Cellular localization

    Nucleus. Colocalizes with ACTN1 and MYOZ2 at the Z line in heart and skeletal muscle.
  • Information by UniProt
  • Alternative names

    • Calmodulin-dependent calcineurin A subunit alpha isoform
    • CALN
    • CALNA
    • CALNA1
    • CAM-PRP catalytic subunit
    • CCN1
    • CNA1
    • PP2B
    • PP2BA_HUMAN
    • PPP2B
    • Ppp3ca
    • protein phosphatase 3, catalytic subunit, alpha isozyme
    • Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform
    see all

Images

  • Example of standard curve obtained with ab139464.

  • Phosphatase activity from a freshly prepared Mouse brain extract. Prior to gel filtration the extract was diluted 1/1 in lysis buffer containing protease inhibitors. After gel filtration and prior to the phosphatase assay, the extract was diluted 1/25 in lysis buffer. The reaction was incubated for 30 min at 30°C.

  • Purified calcineurin positive control. The Calcineurin was incubated 1 hr at 30°C under various buffer conditions. The results demonstrate that Calcineurin activity is inhibited by the EGTA buffer, but not inhibited by 100 nM or 500 nM concentrations of okadaic acid.

Protocols

References

This product has been referenced in:

  • Zhu ZD  et al. SOCE induced calcium overload regulates autophagy in acute pancreatitis via calcineurin activation. Cell Death Dis 9:50 (2018). Read more (PubMed: 29352220) »
See 1 Publication for this product

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