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220487 Chk2, His•Tag®, Human, Recombinant, S. frugiperda

220487
  
Purchase on Sigma-Aldrich

Overview

Replacement Information
Description
Overview

This product has been discontinued.





Recombinant, human checkpoint homologue kinase 2 (Chk2) fused to a His•Tag® sequence at the N-terminus and expressed in S. frugiperda insect cells. Chk2 is involved in sensing DNA damage in eukaryotic cells. It plays an important role in the control of the cell cycle by blocking replication.
Catalogue Number220487
Brand Family Calbiochem®
SynonymsCHK2 checkpoint homolog, CDS1, RAD53, βA44G7, CHEK2
Application Data
Increasing amounts of Chk2, His•Tag®, Active, Human, Recombinant, S. frugiperda were assayed for activity as outlined above.
References
ReferencesBartek, J., et al. 2001. Nat. Rev. Mol. Cell Biol. 12, 877.
Product Information
Unit of DefinitionOne unit is defined as the amount of enzyme required to transfer 1 picomol phosphate to the synthetic peptide substrate KKKVSRSGLYRSPSMPENLNRPR per min at 30°C.
FormLiquid
FormulationIn 270 mM sucrose, 150 mM NaCl, 50 mM Tris-HCl, 1 mM DTT, 0.1 mM EGTA, pH 7.5.
Applications
Biological Information
Purity≥ 95% by SDS PAGE
Specific Activity≥ 25,000 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

Documentation

Chk2, His•Tag®, Human, Recombinant, S. frugiperda Certificates of Analysis

TitleLot Number
220487

References

Reference overview
Bartek, J., et al. 2001. Nat. Rev. Mol. Cell Biol. 12, 877.
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.

Revision22-August-2007 JSW
SynonymsCHK2 checkpoint homolog, CDS1, RAD53, βA44G7, CHEK2
Application Data
Increasing amounts of Chk2, His•Tag®, Active, Human, Recombinant, S. frugiperda were assayed for activity as outlined above.
DescriptionRecombinant, human checkpoint homologue kinase 2 (Chk2) fused to a His•Tag® sequence at the N-terminus and expressed in S. frugiperda insect cells. Chk2 is involved in sensing DNA damage in eukaryotic cells. It plays an important role in the control of the cell cycle by blocking replication.
FormLiquid
FormulationIn 270 mM sucrose, 150 mM NaCl, 50 mM Tris-HCl, 1 mM DTT, 0.1 mM EGTA, pH 7.5.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
Kinase Activity Assay Materials Required • Reaction Buffer: 40 mM MOPS, 2 mM EDTA, pH 7.5 • Substrate: 200 µM CHK peptide substrate, KKKVSRSGLYRSPSMPENLNRPR • Kinase Dilution Buffer: 20 mM MOPS, 10 mM DTT, 1 mM EDTA, 1 mg/ml BSA, 5% glycerol, pH 7.5 • Chk2 Kinase: Chk2, His•Tag®, Active, Human, Recombinant, S. frugiperda diluted to 50-100 ng/3 µl in Kinase Dilution Buffer • Magnesium/ATP Cocktail: 37.5 mM MgCl2, 250 µM ATP • Diluted [γ-32P]ATP: 197 µl Magnesium/ATP Cocktail with 3 µl (30 µCi) [γ-32P]ATP (3000 Ci/mmol) • 0.75% Phosphoric Acid • Acetone Protocol 1. Mix the following components in the specified order in a microcentrifuge tube on ice: • 4 µl Reaction Buffer • 3 µl Substrate • 3 µl Chk2 Kinase (50-100 ng/assay) • 10 µl Diluted [γ-32P]ATP 2. Incubate at 30°C for 10 min. 3. Stop the reaction by placing the tubes on ice. 4. Remove 10 µl of the reaction and spot on P81 paper; allow to bind for 30 s. 5. Immerse the paper in 0.75% Phosphoric Acid and shake gently on a rotator. Discard the Phosphoric Acid and repeat for a total of 3 washes. 6. Wash the paper with acetone. 7. Dry using infrared light. 8. Read the results using a scintillation counter.
Purity≥ 95% by SDS PAGE
Specific activity≥ 25,000 units/mg protein
Unit definitionOne unit is defined as the amount of enzyme required to transfer 1 picomol phosphate to the synthetic peptide substrate KKKVSRSGLYRSPSMPENLNRPR per min at 30°C.
Storage ≤ -70°C
Avoid freeze/thaw
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesBartek, J., et al. 2001. Nat. Rev. Mol. Cell Biol. 12, 877.