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

539672 PKCδ, Human, Recombinant, S. frugiperda

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

Synonyms: Protein Kinase Cδ

539672
Purchase on Sigma-Aldrich

Přehled

Replacement Information

Products

Katalogové čísloBalení ks/bal.
539672-5UG Plastová ampulka 5 μg
Description
OverviewFull-length, recombinant, human PKCδ expressed in S. frugiperda insect cells using a baculovirus expression system. Suitable for studies of PKCδ and its regulation. PKCδ is a Ca2+-independent and phospholipid-dependent enzyme.
Catalogue Number539672
Brand Family Calbiochem®
SynonymsProtein Kinase Cδ
References
ReferencesHug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme that will transfer 1 nmol of phosphate to PKC<sub>ε</sub> Peptide Substrate (<a href ="/Products/ProductDisplay.asp→catNO=539562">Cat. No. 539562</a>) per min at 30°C, pH 7.5.
FormLiquid
FormulationIn 250 mM NaCl, 20 mM HEPES, 5 mM DTT, 2 mM EDTA, 2 mM EGTA, 50% glycerol, 0.05% TRITON® X-100 Detergent, pH 7.5.
Quality LevelMQ100
Applications
Biological Information
Purity≥95% by SDS-PAGE
Specific Activity≥800 units/mg protein
Concentration Label Please refer to vial label for lot-specific concentration
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Standard Handling
Storage ≤ -70°C
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalogové číslo GTIN
539672-5UG 07790788051358

Documentation

PKCδ, Human, Recombinant, S. frugiperda MSDS

Title

Safety Data Sheet (SDS) 

PKCδ, Human, Recombinant, S. frugiperda Certificates of Analysis

TitleLot Number
539672

References

Přehled odkazů
Hug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.
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.

Revision08-September-2008 RFH
SynonymsProtein Kinase Cδ
DescriptionFull-length, recombinant, human PKCδ expressed in S. frugiperda insect cells using a baculovirus expression system. PKC δ is classified as a novel PKC isozyme because it does not exhibit calcium dependence and it is activated in vitro by phorbol esters. Suitable for studies of PKCδ and its regulation. PKCδ is a Ca2+-independent and phospholipid-dependent enzyme.
FormLiquid
FormulationIn 250 mM NaCl, 20 mM HEPES, 5 mM DTT, 2 mM EDTA, 2 mM EGTA, 50% glycerol, 0.05% TRITON® X-100 Detergent, pH 7.5.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
Protocol for Recombinant PKC δ Note: this preparation is purified to near homogeneity, so it may behave differently than crude preparations. In particular, the concentration and quality of the phospholipids used does not seem to be important. Lipid Mixture: 1. Determine the total amount of lipid mixture needed. Each reaction requires 20 µg of phosphatidylserine (1.2 µl of 10 mg/ml phosphatidylserine in CHCl3) and 2 mg of diacylglycerol (0.6 µl of 2 mg/ml diacylglycerol in CHCl3). Preparing 10% more lipid mixture than required is recommended to account for losses while pipetting. 2. Using a Hamilton syringe that has been cleaned with methanol, transfer the required volume of each lipid stock solution to a 12 x 75 mm glass tube. 3. Evaporate off the chloroform. 4. Resuspend in 10 µl of 10 mM HEPES, pH 7.4, 0.3% TRITON® X-100 detergent. 5. Vortex for at least 2 min. 6. Place the lipid mixture in a 40°C water bath for 5 min prior to adding the reaction mixture. Reaction Mixture: 2.4 µl 500 mM HEPES, pH 7.4 6 µl 100 mM MgCl2 6 µl 1 mM EGTA 6 µl 1 mg/ml PKC ε peptide (Cat. No. 539562) 0.6 µl 10 mM ATP 6 µl lipid mixture 0.1 µl γ-ATP (5 mCi/ml) 32.9 µl distilled H2O Total volume should be 60 µl. Prepare a sufficient reaction mixture for all assays to be performed. Reaction: 1. Dispense 60 µl of reaction mixture into each tube and incubate at 30°C. 2. Dilute the PKC δ stock solution (500 ng/µl) to between 20-50 ng/µl in 10 mM Tris-HCl, pH 7.5, 5 mM DTT, and 0.01% TRITON®: X-100 detergent. 3. Add 2 µl of diluted enzyme to each tube. 4. Incubate for 10 min. 5. Stop the reaction by adding 6 µl of 5% phosphoric acid. 6. Incubate on ice for 5 min. 7. Spot 55 µl from each tube to phosphocellulose membranes and allow to dry. 8. Wash the membranes 3X with 5 ml of 0.5% phosphoric acid per filter. 9. Transfer membranes to a scintillation vial and count. Note: This protocol is meant to serve as a guideline and may need to be modified for specific applications.
Purity≥95% by SDS-PAGE
Specific activity≥800 units/mg protein
Unit definitionOne unit is defined as the amount of enzyme that will transfer 1 nmol of phosphate to PKCε Peptide Substrate (Cat. No. 539562) per min at 30°C, pH 7.5.
Storage Avoid freeze/thaw
≤ -70°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesHug, J. and Sarre, T.F. 1993. Biochem. J. 291, 329.
Kazanietz, M.G., et al. 1993. Mol. Pharmacol. 44, 298.
Koide, H., et al. 1992. Proc. Natl. Acad. Sci. USA 89, 1149.