2012 Promega catalogue

Life
Science
Catalog
2012
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Table of
Contents
Cell Signaling
64
PepTag ® Non-Radioactive Protein Kinase
Assays
Product Size Cat.# Price ($)
PepTag ® Non-Radioactive PKC Assay 120 reactions V5330 488.00
PepTag ® Non-Radioactive cAMP-Dependent
Protein Kinase Assay 120 reactions V5340 488.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The PepTag ® Non-Radioactive Protein Kinase Assay Systems
provide a rapid, sensitive and non-radioactive method to detect either Protein
Kinase C (PKC) or Protein Kinase A (PKA) activity. The PepTag ® Assays use
brightly colored, fluorescent peptide substrates that are highly specific for
PKC (PepTag ® C1 Peptide-PLSRTLSVAAK) and PKA (PepTag ® A1 Peptide-
LRRASLG). Phosphorylation of the peptide alters the net charge from +1 to –1.
This change in the net charge allows the phosphorylated and nonphosphorylated
versions of the substrate to be rapidly separated on an agarose gel at
neutral pH. Using fluorescent detection, less than 2ng of purified kinase can
be detected in less than 2 hours. The PepTag ® Non-Radioactive Protein Kinase
Assay Systems can detect kinase activity in partially purified samples as well as
purified preparations of enzymes, making it a good choice for the rapid screening
of column fractions or the screening of kinase activators and inhibitors. In
addition to the assay components, each system includes purified kinase for use
as a positive control.
Features:
• Nonradioactive: The fluorescent tag on the peptide substrate facilitates
quantitation of the phosphorylation reaction without the use of radioactivity.
• Low Background: Phosphorylation of the colored peptide supplied with
the system is used to measure kinase activity; therefore phosphorylation
of other substrates occurring naturally in the sample does not add to the
kinase activity measured.
• Convenient: Measure the phosphorylated peptide using a densitometer,
spectrophotometer, 96-well plate reader, or fluorometer.
Storage Conditions: Store at –70°C.
Protocol Part#
PepTag ® Assay for Non-Radioactive Detection of Protein Kinase C or
cAMP-Dependent Protein Kinase Technical Bulletin
TB132
nonphosphorylated
(+1 net charge)
colored peptide substrate
(+1 net charge)
Incubate peptide with
kinase-containing sample.
phosphorylated
(–1 net charge)
Separate phosphorylated and nonphosphorylated
forms by agarose gel electrophoresis.
Net Charge
of Peptide
+1
0
–1
anode
cathode
P =
Peptide Species
nonphosphorylated
P =
phosphorylated
Schematic diagram of PepTag ® Non-Radioactive Protein Kinase Assay
procedure.
Protein Kinases
0383MB
Protein Kinases
cAMP-Dependent Protein Kinase, Catalytic
Subunit
Product
cAMP-Dependent Protein Kinase,
Size Conc. Cat.# Price ($)
Catalytic Subunit 2,500 u 1.5–3 mg/ml V5161 323.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The purified 40kDa cAMP-Dependent Protein Kinase (PKA),
Catalytic Subunit, may be used to phosphorylate target proteins or for in vitro
enzymological studies of neural and hormonal signal transduction. Intracellular
targets include ion channels, transcriptional activator proteins, and regulatory
enzymes of glycogen metabolism.
Features:
• Highly Pure: The PKA Catalytic Subunit is purified from a recombinant E.
coli strain expressing the catalytic subunit of bovine PKA and is 90% pure.
Storage Conditions: Store at –70°C.
Protocol Part#
cAMP-Dependent Protein Kinase, Catalytic Subunit Product
Information
9PIV516
DNA-Dependent Protein Kinase
Product Size Cat.# Price ($)
DNA-Dependent Protein Kinase
For Research Use Only. Not for Use in Diagnostic Procedures.
2,500 u V5811 352.00
Description: DNA-Dependent Protein Kinase (DNA-PK) phosphorylates
several DNA-binding substrates in vitro, including the tumor suppressor protein
p53, the SV40 large T antigen and several transcription factors. DNA-PK is
thought to play a role in controlling gene regulation and cell growth.
DNA-PK is isolated from HeLa nuclear extracts as a complex consisting of a
400kDa catalytic subunit and a 155kDa heterodimeric DNA-binding component
named Ku, which itself consists of subunits of approximately 85kDa and
70kDa.
Storage Conditions: Store at –70°C.
Protocol Part#
DNA-Dependent Protein Kinase Product Information 9PIV581
cGMP-Dependent Protein Kinase
Product Size Cat.# Price ($)
cGMP-Dependent Protein Kinase (α-Isozyme)
For Research Use Only. Not for Use in Diagnostic Procedures.
6,000 u V5171 352.00
Description: cGMP-Dependent Protein Kinase is a serine/threonine protein
kinase present in smooth muscle and a variety of other tissues. The kinase is
a 78kDa polypeptide composed of a regulatory domain and a catalytic domain
and is active as a homodimer.
Specific Activity: >1,000u/μg (kinase activity).
Features:
• Highly Pure: cGMP-Dependent Protein Kinase has been purified by the
method of Corbin and Doskeland and is >90% pure as determined by
SDS-PAGE (single band).
Storage Conditions: Store at –70°C.
Protocol Part#
cGMP-Dependent Protein Kinase (Alpha Isozyme) Product
Information
9PIV517
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