2012 Promega catalogue

Cell Signaling
Protein Kinase Substrates
Product Size Conc. Cat.# Price ($)
Kemptide (PKA) Peptide Substrate 1 mg 10 mg/ml V5601 64.00
Neurogranin (28–43) (PKC) Peptide
Substrate 1 mg 10 mg/ml V5611 275.00
DNA-Dependent Protein Kinase Peptide
Substrate 1 mg 10 mg/ml V5671 427.00
cdc2 Protein Kinase Peptide Substrate
cGMP-Dependent Protein Kinase Peptide
1 mg 10 mg/ml V2211 432.00
Substrate 1 mg 10 mg/ml V7451 432.00
Casein Kinase I Peptide Substrate 1 mg 10 mg/ml V7441 267.00
Casein Kinase II Peptide Substrate 1 mg 10 mg/ml V5661 297.00
Protein Phosphatases and
Phosphatase Assays
PPase-2A
Product Size Cat.# Price ($)
PPase-2A
For Research Use Only. Not for Use in Diagnostic Procedures.
25 u V6311 662.00
Description: Protein Phosphatase-2A (PPase-2A) is a serine/threonine phosphatase
isolated from human red blood cells. It is isolated as the heterodimer
of 60kDa (A) and 36kDa (C) subunits. It has the ability to dephosphorylate
the α-subunit of phosphorylase kinase. With its 36–38kDa catalytic subunit,
PPase-2A has broad substrate specificity and may play a regulatory role in DNA
replication, transcription, protein synthesis, mitosis and glycogen metabolism.
PPase-2A is stimulated in vitro by basic proteins such as protamine, histones
and polylysine. The enzyme is inhibited by several environmental toxins and
tumor promoters such as okadaic acid and microcystin-LR. The chemically
synthesized phosphopeptide, RRA(pT)VA (available in the Ser/Thr Phosphatase
Assay System, Cat.# V2460), is an excellent substrate for PPase-2A.
Storage Conditions: Store at –20°C.
Protocol Part#
PPase-2A Product Information 9PIV631
PPase-2B
Product Size Cat.# Price ($)
PPase-2B
For Research Use Only. Not for Use in Diagnostic Procedures.
10 u V6361 186.00
Description: PPase-2B is a heterodimeric enzyme composed of a 19kDa
calcium-binding subunit and a catalytic subunit (61kDa) that binds calmodulin.
PPase-2B was originally identified based on its calcium- and calmodulindependent
activity toward phosphorylase kinase and inhibitor-1. PPase-2B is
identical to the brain protein calcineurin, which constitutes up to 1% of total
brain protein. The immunosuppressive drugs FK-506 and cyclosporin A inhibit
PPase-2B activity in immune cells, implicating this enzyme in regulation of
the immune system. PPase-2B also plays a major role in regulating secretory
functions of a variety of cells.
PPase-2B is less sensitive to okadaic acid than PPase-2A and PPase-1,
requiring micromolar concentrations of okadaic acid for inhibition. It is not
inhibited by Inhibitor-1 or Inhibitor-2. Promega PPase-2B is isolated from
bovine brain.
Storage Conditions: Store at –70°C.
Protocol Part#
PPase-2B Product Information 9PIV636
For complete and up-to-date product information visit: www.promega.com/catalog
Non-Radioactive Phosphatase Assay Systems
Product Size Cat.# Price ($)
Serine/Threonine Phosphatase Assay System 96 reactions V2460 616.00
Tyrosine Phosphatase Assay System 96 reactions V2471 616.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The Non-Radioactive Phosphatase Assay Systems are convenient
and flexible alternative for measuring protein phosphatase activity. These
systems measure the absorbance of a molybdate:malachite green:phosphate
complex to determine the amount of free phosphate generated in a reaction.
These systems work with many buffer conditions and substrates, including
naturally phosphorylated proteins or synthetic phosphopeptides. The Serine/
Threonine Phosphatase Assay System contains the chemically synthesized
phosphopetide, RRA(pT)VA, a peptide substrate compatible with several serine/
threonine phosphatases such as the Protein Phosphatases 2A, 2B, and 2C.
However the supplied phosphopeptide is a poor substrate for Protein
Phosphatase 1 because of its more stringent structural requirements.
The Tyrosine Phosphatase Assay System contains two chemically synthesized
phosphopeptides, END(pY)INASL and DADE(pY)LIPQQG, that serve as
substrates for many protein tyrosine phosphatases. The effective range for the
detection of phosphate released during an assay using the Phosphatase Assay
Systems is 100–4,000pmol of phosphate. The Phosphatase Assay Systems
can measure phosphatase activity in crude cell or tissue extracts or in partially
fractionated and purified samples. For application in crude extracts, the high
concentration of phosphate is eliminated before performing the assay using the
supplied Spin Columns, which rapidly and effectively remove free phosphate
and other low-molecular-weight inhibitors from the sample. In addition, a
unique Molybdate Dye Additive, which is combined with the Molybdate Dye
Solution aids in the solubilization of proteins exposed to the acid conditions of
the Molybdate Dye Solution, which alone could potentially cause precipitation
of the proteins.
Storage Conditions: Store the entire kit at 4°C.
Protocol Part#
Serine/Threonine Phosphatase Assay System Technical Bulletin TB218
Tyrosine Phosphatase Assay System TB212
pmol phosphate/min/µg protein
4
3
2
1
0
S = 100µM provided substrate, no inhibitors
F = 50mM Sodium Fluoride
V = 1mM Sodium Vanadate
nM OA = 12nM Okadaic Acid
µM OA = 5µM Okadaic Acid
I = 50ng/ml PPase-1 Inhibitor-1
S
F
V
nM OA
µM OA
I
S
nM OA
S
nM OA
µM OA
PPase-2A PPase-2B PPase-2C
Buffer
Buffer Buffer
Serine/Threonine phosphatase activity in HeLa cell extract using the
Serine/Threonine Phosphatase Assay System.
0960MA02_5B
69
3
Cell Signaling
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