2012 Promega catalogue

Cell Signaling
SignaTECT ® Protein Kinase Assay Systems
Product Size Cat.# Price ($)
SignaTECT ® cAMP-Dependent Protein Kinase
(PKA) Assay System 96 reactions V7480 567.00
SignaTECT ® Protein Kinase C (PKC) Assay
System 96 reactions V7470 567.00
SignaTECT ® Protein Tyrosine Kinase (PTK)
Assay System 96 reactions V6480 686.00
SignaTECT ® Calcium/Calmodulin-Dependent
Protein Kinase (CaM KII) Assay System 96 reactions V8161 567.00
SignaTECT ® DNA-Dependent Protein Kinase
Assay System 96 reactions V7870 567.00
SignaTECT ® cdc2 Protein Kinase Assay
System 96 reactions V6430 567.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The SignaTECT ® Protein Kinase Assay Systems contain the
proprietary SAM 2® Biotin Capture Membrane. The streptavidin-coated SAM 2®
Membranes possess high binding capacity and high specificity characteristics,
which 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. The SAM 2® Membrane format does not
require as much “hands-on” manipulation as other methods used to measure
kinase activity. Following the kinase reaction, samples are spotted onto the
SAM 2® Membrane, and a series of short wash steps are performed to remove
nonspecific label. The process is complete in less than 1 hour. In addition, the
nature of the SAM 2® Membrane allows it to be used under a variety of buffer
and reaction conditions (e.g., cell extracts), which many other methods do not
allow. Last, the high binding capacity allows use of the SignaTECT ® Systems
for kinetic studies.
Each SignaTect ® system contains specific biotinylated peptide substrates for
the appropriate kinase as well as the necessary reaction components. The
researcher must supply [γ- 32 P]ATP.
Storage Conditions: Store all SignaTECT ® Systems except V7470 at –20°C.
Store Cat.# V7470 at –70°C.
Protocol Part#
SignaTECT ® cAMP-Dependent Protein Kinase (PKA) Assay System
Technical Bulletin
TB241
SignaTECT ® Protein Kinase C (PKC) Assay System Technical Bulletin TB242
SignaTECT ® Protein Tyrosine Kinase Assay System Technical Bulletin TB211
SignaTECT ® Calcium/Calmodulin-Dependent Protein Kinase Assay
System Technical Bulletin
TB279
SignaTECT ® DNA-Dependent Protein Kinase Assay System Technical
Bulletin
TB250
SignaTECT ® cdc2 Protein Kinase Assay System Technical Bulletin TB227
For complete and up-to-date product information visit: www.promega.com/catalog
Biotin–(C) 6 –XXXX(Y/S/T)XXXX
Biotinylated Peptide Substrate
+
Protein Kinase Sample
+
Reaction Buffer
+
[γ-32P]ATP Combine peptide substrate,
[γ-32P]ATP, buffer
and sample. Incubate
at 30°C for 2–10 minutes.
SAM 2® Biotin
Capture
Membrane
Reaction
Components
PO4 Biotin–(C) 6 –XXXX(Y/S/T)XXXX
Terminate the kinase
reaction with
Termination Buffer.
Capture the biotinylated
peptide substrate by
spotting reactions on
the Membrane.
PO4 Biotin–(C) 6 –XXXX(Y/S/T)XXXX
Wash Membrane to
remove unbound
reaction components.
Quantitate
• Scintillation counter
• Phosphorimaging system
• Autoradiography
Schematic diagram of the SignaTECT ® Protein Kinase Assay protocol.
Protocol steps to prepare, run and analyze a specific protein kinase
activity using any of the SignaTECT ® Protein Kinase Assay Systems.
1542MA05_7A
63
3
Cell Signaling
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