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324881 PERK, GST-Fusion, Human, Recombinant, E. coli

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

Synonyms: EIF2AK3 PKR

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Overview

Replacement Information
Description
Overview

This product has been discontinued.



Recombinant, human PERK consisting of amino acids 563-1115 fused at the N-terminus to GST and expressed in E. coli. PERK phosphorylates the α-subunit of eukaryotic translation initiation factor 2 (EIF2α), which leads to its inactivation and reduces translation initiation. The active kinase is useful for the study of PERK regulation and for inhibitor screening.
Note: the calculated MW is 88,000, but the protein runs at ~115,000 on SDS-PAGE.
Catalogue Number324881
Brand Family Calbiochem®
SynonymsEIF2AK3 PKR
Application Data
The activity of PERK was measured as outlined in the Recommended Reaction Conditions.
References
ReferencesBaltzis, D., et al. 2007. J Biol Chem. 43, 31675.
Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.
Product Information
Unit of DefinitionKinase activity is measured as the molar amount of phosphate incorporated into SMAD3 at 30°C using a final concentration of 50 µM ATP.
FormLiquid
FormulationIn 50 mM Tris-HCl, 50 mM NaCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
Quality LevelMQ100
Applications
Biological Information
Purity≥90% by SDS-PAGE
Specific ActivityPlease refer to the lot-specific Certificate of Analysis
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
Catalogue Number GTIN
324881 0

Documentation

PERK, GST-Fusion, Human, Recombinant, E. coli SDS

Title

Safety Data Sheet (SDS) 

PERK, GST-Fusion, Human, Recombinant, E. coli Certificates of Analysis

TitleLot Number
324881

References

Reference overview
Baltzis, D., et al. 2007. J Biol Chem. 43, 31675.
Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.
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.

Revision06-September-2017 JSW
SynonymsEIF2AK3 PKR
Application Data
The activity of PERK was measured as outlined in the Recommended Reaction Conditions.
DescriptionRecombinant, human PERK consisting of amino acids 563-1115 fused at the N-terminus to GST and expressed in E. coli. PERK phosphorylates the α-subunit of eukaryotic translation initiation factor 2 (EIF2α), which leads to its inactivation and reduces translation initiation. The active kinase is useful for the study of PERK regulation and for inhibitor screening. Note: the calculated MW is 88,000, but the protein runs at ~115,000 on SDS-PAGE.
FormLiquid
FormulationIn 50 mM Tris-HCl, 50 mM NaCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
Kinase assay conditions: Dilute purified PERK in kinase dilution buffer (5 mM MOPS, pH 7.2, 4 mM MgCl2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 400 µM EDTA, 50 ng/µl BSA, 5% glycerol; add DTT to 50 µM just prior to use) to the desired concentration. Mix 10 µl diluted enzyme, 5 µl SMAD3 substrate (200 ng/ml stock), and 5 µl H2O and incubate for 5 min at 30°C. Add 5 µl ATP mixture [(150 µl of 10 mM ATP, 100 µl of 1 mCi/100µl [32P]ATP, and 5.75 ml kinase assay buffer (25 mM MOPS, pH 7.2, 25 mM MgCL2, 12.5 mM β-glycerophosphate, 5 mM EGTA, 2 mM EDTA with 250 µM DTT just prior to use)] and incubate for 15 min at 30°C. Terminate the reaction by spotting 20 µl of the reaction mixture onto a phosphocellulose P81 paper, dry, and wash several times with 1% phosphoric acid prior to counting in the presence of scintillation fluid.
Purity≥90% by SDS-PAGE
Specific activityPlease refer to the lot-specific Certificate of Analysis
Unit definitionKinase activity is measured as the molar amount of phosphate incorporated into SMAD3 at 30°C using a final concentration of 50 µM ATP.
Storage Avoid freeze/thaw
≤ -70°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesBaltzis, D., et al. 2007. J Biol Chem. 43, 31675.
Koumenis, C., et al. 2002. Mol Cell Biol. 21, 7405.