SignaTECT® Protein Kinase Assay Systems

V8161_SignaTECT--CaMKII-Protein-Kinase-Assay_3
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Radioactive Assays for Detection of Kinase Activity

  • Assays for calcium/calmodulin-dependent protein kinase

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Catalog number selected: V8161

$ 630.00
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SignaTECT® Protein Kinase Assay Systems
96 reactions
$ 630.00
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Convenient, 1-Hour Kinase Activity Assays

The SignaTECT® Protein Kinase Assay Systems provide a rapid, radioactivity-based method for detection of kinase activity. The method is based on capture of phosphorylated labeled substrates on the SAM2® Biotin Capture Membrane, which offers low background and high signal-to-noise ratios, even for complex samples such as crude cell extracts, while retaining the high substrate capacity necessary to maintain optimum reaction kinetics.

Each SignaTECT® System contains a highly specific biotinylated peptide substrate for the appropriate kinase as well as the necessary reaction components. The researcher must supply [γ-32P]ATP.

Method Overview

After the kinase reaction is complete, the biotinylated, 32P-labeled substrate is recovered from the reaction mix using the SAM2® Biotin Capture Membrane. After samples are spotted onto the SAM2® Membrane a series of short washes are performed to remove nonspecific label. The process is complete in less than 1 hour. The SAM2® Membrane is prenumbered and partially cut so that individual squares can be easily identified, separated and placed in scintillation vials. Alternatively, the intact SAM2® Membrane can be analyzed by phosphorimaging or conventional autoradiography.

Advantages of the SAM2® Membranes

  • High binding capacity and high specificity characteristics produce lower backgrounds and higher signal-to-noise ratios compared to the traditional P81 phosphocellulose method of capture and measurement.
  • The perforated and numbered membrane allows researchers to measure from 1 up to 96 kinase reactions and does not require as much "hands-on" manipulation as other methods used to measure kinase activity.
  • Can be used under a variety of buffer/reaction conditions (e.g., cell extracts), which many other methods do not allow.
  • High binding capacity allows use of the SignaTECT® Systems for kinetic studies.

Specifications

What's in the box?

Item Part # Size Concentration

ATP

V222A 1 × 600μl 0.5mM

BSA

V275A 1 × 0.2ml 10mg/ml

SAM2® Biotin Capture Membrane

V2861 1 × 96 samples

Termination Buffer

V594A 2 × 700μl

CaM KII Biotinylated Peptide Substrate

V817A 1 × 300μl

CaM KII Reaction 5X Buffer

V818A 1 × 1ml

CaM KII Control 5X Buffer

V819A 1 × 500μl

CaM KII Activation 5X Buffer

V820A 1 × 500μl

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

U.S. Pat. Nos. 6,066,462, 6,348,310 and 6,753,157.

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