PTP1B drug discovery kit



BML-AK822-0001	1 Kit	397.00 USD

The PTP1B Tyrosine Phosphatase Drug Discovery Kit is a colorimetric, non-radioactive assay designed to measure the phosphatase activity of purified PTP1B. This 96-well assay is useful for screening inhibitors and modulators of PTP1B activity.The kit includes human, recombinant PTP1B (residues 1-322; MW=37.4 kDa), expressed in E. coli. The detection of free phosphate released is based on the classic Malachite green assay and offers the advantages of convenient, 1-step detection and excellent sensitivity, without radioactivity. PTP1B (protein tyrosine phosphatase-1B) is a ubiquitous, nontransmembrane protein tyrosine phosphatase, originally identified in human placenta.

Quantity:	96 assays

Kit/Set Contains:	PTP 1B Enzyme (human recombinant) (Prod. No. BML-SE332-9092)(5 µg; 0.1 µg/µl in 50 mM HEPES, pH 7.2, 1 mM EDTA, 1 mM DTT, 0.05% NP-40, 10% (v/v) glycerol)Storage: -70°C. Avoid freeze/thaw cycles Substrate (“IR5” Insulin Receptor ß, residues 1142-1153, pY¹¹⁴⁶) (Prod. No. BML-P315-9090)(1 mg; Lyophilized solid)Storage: -20 or -70°C. 2X Assay Buffer (Prod. No. BML-KI467)(20 ml; 100 mM MES, pH 6.0, 300 mM NaCl, 2 mM EDTA, 2 mM DTT, 0.1% NP-40 Storage: -70°C Biomol Red™ Concentrated Phosphate Detection Reagent (Prod. No. BML-KI468)(5 ml; Liquid in screw-cap plastic bottle)Storage: 4°C. Long-term at -70°C. Suramin (PTP1B inhibitor) (Prod. No. BML-KI469)(10 mg; Solid)Storage: -70°C Phosphate Standard (Prod. No. BML-KI470)(0.5 ml; 100 µM in 1X Assay Buffer)Storage: -20 or -70°C ½-VOLUME MICROPLATE (Prod. No. BML-KI101)Storage: RT

Long Term Storage:	-80°C

Use/Stability:	Store all components except the microtiter plate at -70°C for the highest stability. The PTP1B enzyme component BML-SE332-9092 must be handled particularly carefully in order to retain maximal enzymatic activity. Thaw it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused enzyme should be ‘snap’ frozen, e.g. in liquid nitrogen or a dryice/ethanol bath, and stored at -70°C. If only a few assays are to be performed each day, the PTP1B may be divided into several aliquots (best if ≥10 µl)) to help minimize freeze/thaw cycles.

Background / Technical Information:	Please click here for the comprehensive product datasheet.

General Literature References

Protein-tyrosine phosphatase 1B deficiency reduces insulin resistance and the diabetic phenotype in mice with polygenic insulin resistance: B. Xue et al.; J. Biol. Chem. 282, 23829 (2007), Abstract;

SIRT1 improves insulin sensitivity under insulin-resistant conditions by repressing PTP1B: C. Sun et al.; Cell. Metab. 6, 307 (2007), Abstract;

Suramin derivatives as inhibitors and activators of protein-tyrosine phosphatases: D.F. McCain et al.; J. Biol. Chem 279, 14713 (2004), Abstract;

Mechanism of transmembrane signaling: insulin binding and the insulin receptor: F.P. Ottensmeyer et al.; Biochemistry 39, 12103 (2000), Abstract;

Molecular basis for the dephosphorylation of the activation segment of the insulin receptor by protein tyrosine phosphatase 1B: A. Salmeen et al.; Mol. Cell. 6, 1401 (2000), Abstract;

Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene: M. Elchbly et al.; Science 283, 1544 (1999), Abstract;

Crystal structure of the activated insulin receptor tyrosine kinase in complex with peptide substrate and ATP analog: S.R. Hubbard; Embo. J. 16, 5572 (1997), Abstract;

Identification of a second aryl phosphate-binding site in protein-tyrosine phosphatase 1B: a paradigm for inhibitor design: Y.A. Pius et al.; PNAS 94, 13420 (1997), Abstract;

Direct binding of the proline-rich region of protein tyrosine phosphatase 1B to the Src homology 3 domain of p130(Cas): F. Liu et al.; J. Biol. Chem. 271, 31290 (1996), Abstract;

Characterization and kinetic analysis of the intracellular domain of human protein tyrosine phosphatase beta (HPTP beta) using synthetic phosphopeptides: K.W. Harder et al.; Biochem. J. 298, 395 (1994), Abstract;

Sequential dephosphorylation of a multiply phosphorylated insulin receptor peptide by protein tyrosine phosphatases: C. Ramachandran et al.; Biochemistry 31, 4232 (1992), Abstract;

Use of fluorinated tyrosine phosphates to probe the substrate specificity of the low molecular weight phosphatase activity of calcineurin: B. Martin et al.; J. Biol. Chem. 260, 14932 (1985), Abstract;