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.
It is implicated in the negative regulation of insulin receptor signaling, and is a potential therapeutic target for treatment of type 2 diabetes and obesity. The phosphopeptide substrate supplied with this kit ("IR5", Prod. No. BML-P315, pTyr1158 (a.k.a. pTyr1146, mature peptide numbering)) contains sequence from the insulin receptor β subunit domain that must be autophosphorylated to achieve full receptor kinase activation. This "activation loop" is the target of several protein phosphatase regulators of insulin signaling, including, notably, PTP1B. The PTP1B inhibitor suramin is supplied as a control for inhibitor detection. Suramin is a reversible and competitive inhibitor of PTP1B, with a Ki of 5.5 µM.
Product Details
| Applications: | Colorimetric detection, HTS
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| 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. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: | PTP 1B Enzyme, Substrate (“IR5” Insulin Receptor β, residues 1142-1153, pY1146), Assay Buffer, Biomol Red™ Concentrated Phosphate Detection Reagent, Suramin (PTP1B inhibitor), Phosphate Standard, and Microplate (½-volume) |
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| UniProt ID: | P18031 |
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| Regulatory Status: | RUO - Research Use Only |
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| Compatibility: | This product is compatible with the Absorbance 96 Plate Reader.
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Product Literature References
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Abstract;
Synthesis, biological evaluation and in silico studies of 5-(3-methoxybenzylidene)thiazolidine-2,4-dione analogues as PTP1B inhibitors: M.K. Mahapatra, et al.; Bioorg. Chem.
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Abstract;
Optimization of extraction parameters of PTP1β (protein tyrosine phosphatase 1β), inhibitory polyphenols, and anthocyanins from Zea mays L. using response surface methodology (RSM): S.H. Hwang, et al.; BMC Complement. Altern. Med.
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Application(s): Enzymatic assay measurement of PTP1β inhibitory activity,
Abstract;
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In vitro cytotoxic, in vitro and in vivo antidiabetic activity of roylea cineria: R.P. Bahuguna, et al.; Sci. Revs. Chem. Commun.
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Application(s): Assay using plant fractions,
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Protein-tyrosine phosphatase 1B (PTP1B) is a novel regulator of central brain-derived neurotrophic factor and tropomyosin receptor kinase B (TrkB) signaling: C. Ozek, et al.; J. Biol. Chem.
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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.
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Abstract;
SIRT1 improves insulin sensitivity under insulin-resistant conditions by repressing PTP1B: C. Sun et al.; Cell. Metab.
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Abstract;
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Abstract;
Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene: M. Elchbly et al.; Science
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Crystal structure of the activated insulin receptor tyrosine kinase in complex with peptide substrate and ATP analog: S.R. Hubbard; Embo. J.
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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
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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.
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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.
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Abstract;
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