Cell Signaling Protein Kinase Substrates Product Kemptide (PKA) Peptide Substrate Neurogranin(28–43) (PKC) Peptide Substrate DNA-Dependent Protein Kinase Peptide Substrate cGMP-Dependent Protein Kinase Peptide Substrate Casein Kinase I Peptide Substrate Casein Kinase II Peptide Substrate Size Conc. Cat.# Price (Fr) 1 mg 10 mg/ml V5601 72.00 1 mg 10 mg/ml V5611 248.00 1 mg 10 mg/ml V5671 360.00 cdc2 Protein Kinase Peptide Substrate 1 mg 10 mg/ml V2211 364.00 1 mg 10 mg/ml V7451 404.00 1 mg 10 mg/ml V7441 288.00 1 mg 10 mg/ml V5661 297.00 Protein Phosphatases and Phosphatase Assays PPase-2A Product PPase-2A For Research Use Only. Not for Use in Diagnostic Procedures. 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 PPase-2A Product Information PPase-2B Product PPase-2B For Research Use Only. Not for Use in Diagnostic Procedures. 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 PPase-2B Product Information Part# 9PIV636 For complete and up-to-date product information visit: www.promega.com/catalog 69 Size Cat.# Price (Fr) 10 u V6361 225.00 Part# 9PIV631 Size Cat.# Price (Fr) 25 u V6311 811.00 Product Non-Radioactive Phosphatase Assay Systems Size Cat.# Price (Fr) Serine/Threonine Phosphatase Assay System 96 reactions V2460 644.00 Tyrosine Phosphatase Assay System 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 Serine/Threonine Phosphatase Assay System Technical Bulletin Tyrosine Phosphatase Assay System 0 1 2 3 4 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 Part# TB218 TB212 96 reactions V2471 644.00 3 PPase-2A Buffer PPase-2B Buffer PPase-2C Buffer Serine/Threonine phosphatase activity in HeLa cell extract using the Serine/Threonine Phosphatase Assay System. Cell Signaling pmol phosphate/min/µg protein S F V nM OA µM OA I S nM OA S nM OA µM OA 0960MA02_5B Pagina 72
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