Millipore Sigma Vibrant Logo
Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

207008 Calcineurin Substrate - Calbiochem

An excellent peptide substrate for calcineurin.

Calcineurin Substrate - Calbiochem MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Synonyms: PP2B Substrate, RII Phosphopeptide, DLDVPIPGRFDRRVpSVAAE

View Products on Sigmaaldrich.com
207008
Purchase on Sigma-Aldrich

Overview

Replacement Information

Products

Catalogue NumberPackaging Qty/Pack
207008-500UG Plastic ampoule 500 μg
Description
OverviewAn excellent peptide substrate for calcineurin (Protein Phosphatase 2B, PP2B; Cat. No. 539568). The sequence is derived from the PKA regulatory subunit type II (RII) and is phosphorylated on the Ser residue. Release of phosphate can be quantified colorimetrically using the malachite green method.
Catalogue Number207008
Brand Family Calbiochem®
SynonymsPP2B Substrate, RII Phosphopeptide, DLDVPIPGRFDRRVpSVAAE
References
ReferencesDonella-Deana, A., et al. 1994. Eur. J. Biochem. 219, 109.
Enz, A., et al. 1994. Anal. Biochem. 216, 147.
Harder, K.W., et al. 1994. Biochem. J. 298, 395.
Fruman, D.A., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3686.
Product Information
ATP CompetitiveN
FormLyophilized
FormulationSupplied as a trifluoroacetate salt.
Hill FormulaC₉₂H₁₅₁N₂₈O₃₂P
Chemical formulaC₉₂H₁₅₁N₂₈O₃₂P
Hygroscopic Hygroscopic
ReversibleN
Quality LevelMQ100
Applications
Biological Information
Primary TargetCalcineurin (Protein Phosphatase 2B, PP2B)
Purity≥95% by HPLC
Physicochemical Information
Cell permeableN
Peptide SequenceH-Asp-Leu-Asp-Val-Pro-Ile-Pro-Gly-Arg-Phe-Asp-Arg-Arg-Val-Ser(PO₃H₂)-Val-Ala-Ala-Glu-OH
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Ambient Temperature Only
Toxicity Standard Handling
Storage -20°C
Hygroscopic Hygroscopic
Do not freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze at -20°C. Stock solutions are stable for up to 3 months at -20°C.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
207008-500UG 04055977203233

Documentation

Calcineurin Substrate - Calbiochem SDS

Title

Safety Data Sheet (SDS) 

Calcineurin Substrate - Calbiochem Certificates of Analysis

TitleLot Number
207008

References

Reference overview
Donella-Deana, A., et al. 1994. Eur. J. Biochem. 219, 109.
Enz, A., et al. 1994. Anal. Biochem. 216, 147.
Harder, K.W., et al. 1994. Biochem. J. 298, 395.
Fruman, D.A., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3686.

Brochure

Title
Protein Phosphatases Technical Bulletin
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision27-May-2008 RFH
SynonymsPP2B Substrate, RII Phosphopeptide, DLDVPIPGRFDRRVpSVAAE
DescriptionAn excellent peptide substrate for calcineurin (Protein Phosphatase 2B, PP2B, Cat. No. 539568). The sequence is derived from the PKA regulatory subunit type II (RII) and is phosphorylated on the Ser residue. Release of phosphate can be quantified colorimetrically using malachite green method.
FormLyophilized
FormulationSupplied as a trifluoroacetate salt.
Recommended reaction conditions
Malachite Green Assay Note: This protocol is provided only as a general guide. Researchers should standardize this assay for their own specific needs and should consult published literature. Reagents • 4.2% Ammonium molybdate: prepare a 4.2% (w/v) solution of ammonium molybdate in 4 M HCl • Malachite Green: prepare a 0.045% (w/v) solution of malachite green • Malachite Green Solution: Mix 1 volume of 4.2% ammonium molybdate (prepared as above) with 3 volumes of 0.045% malachite green (as prepared above); stir for a minimum of 30 min; filter sterilize using a 0.22 µm filter. This solution is stable at 4°C for 6 months. • Malachite Green/Tween Solution: 0.01% Tween®-20 detergent (v/v) is added to an aliquot of Malachite Green Solution just prior to use • 2X assay buffer: 200 mM NaCl, 100 mM Tris, 12 mM MgCl2, 1 mM CaCl2, 1 mM DTT, 0.05% NP-40, pH 7.5. Assay buffer can be stored in aliquots at -70°C. • Calcineurin Substrate: Prepare a 750 µM (1.64 mg/ml) stock solution in H2O • Calmodulin, Bovine Brain (Cat. No. 208690 or 208694): prepare a 25 µM stock solution in H2O. Prepare a working solution of calmodulin by diluting the stock solution 1:50 in 2X assay buffer; a total of 25 µl is needed per reaction. • Cell lysate or purified PP2B (Calcineurin) (Cat. No. 539568) • Phosphatase inhibitors, as desired • 1/2 volume plate • KH2PO4: prepare a phosphate standard stock solution by dissolving in 0.05 N HCl to an appropriate concentration of phosphorous (nmol or pmol/ml). Prepare serial dilutions just prior to use for generation of a standard curve. Use 25 µl of each standard dilution plus 25 µl of 2X assay buffer; add 100 µl of Malachite Green/Tween Solution and measure absorbance (see below). • Plate reader capable of measuring absorbance at 620 nm Protocol • Prepare phosphate standard samples in the desired number of wells in a microtiter plate. It is recommended that the standards be performed in duplicate. • Prepare sample wells: • 25 µl of 2X assay buffer • 10 µl of Calcineurin Substrate • 5 µl of Calcineurin preparation (cell lysate or ~40 U of purified enzyme) Note: control samples may be included by the addition of appropriate phosphatase inhibitors. Adjust the H2O volume appropriately to maintain a 50 µl total reaction volume. • Incubate for the desired time (e.g. 30 min at 30°C) • Terminate the reactions by adding 100 µl of Malachite Green/Tween Solution to each well. Continue incubating for 15-30 min. • Read the absorbance at 620 nm. • Compare samples to the standard curve to determine phosphatase activity.
Chemical formulaC₉₂H₁₅₁N₂₈O₃₂P
Peptide SequenceH-Asp-Leu-Asp-Val-Pro-Ile-Pro-Gly-Arg-Phe-Asp-Arg-Arg-Val-Ser(PO₃H₂)-Val-Ala-Ala-Glu-OH
Purity≥95% by HPLC
SolubilityH₂O (2 mg/ml)
Storage -20°C
Hygroscopic
Do Not Freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze at -20°C. Stock solutions are stable for up to 3 months at -20°C.
Toxicity Standard Handling
ReferencesDonella-Deana, A., et al. 1994. Eur. J. Biochem. 219, 109.
Enz, A., et al. 1994. Anal. Biochem. 216, 147.
Harder, K.W., et al. 1994. Biochem. J. 298, 395.
Fruman, D.A., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 3686.