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Rediplate 96 enzchek tyrosine phosphatase assay kit

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The RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit is a fluorescence-based assay that measures the activity of tyrosine phosphatases. The kit provides a substrate and necessary reagents to quantify tyrosine phosphatase activity in a 96-well plate format.

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Lab products found in correlation

Shp 1 activity assay, by Thermo Fisher Scientific (6 mentions) Protein g sepharose 4 fast flow, by GE Healthcare (5 mentions) Anti shp 1 polyclonal antibody, by Santa Cruz Biotechnology (2 mentions) Protein a g plus agarose, by Santa Cruz Biotechnology (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Protein g agarose beads, by Roche (1 mentions) Scrambled control sirna, by Thermo Fisher Scientific (1 mentions) Imatinib, by Merck Group (1 mentions) Anti shp2, by Santa Cruz Biotechnology (1 mentions) Lipofectamine ltx reagent, by Thermo Fisher Scientific (1 mentions) Anti p stat5, by Cell Signaling Technology (1 mentions) Anti myc tag, by Cell Signaling Technology (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions) Anti beclin 1, by Cell Signaling Technology (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti c cbl, by Cell Signaling Technology (1 mentions) Supersep phos tag, by Fujifilm (1 mentions) Anti stat5, by Cell Signaling Technology (1 mentions) Anti p shp2, by Cell Signaling Technology (1 mentions) Oridonin, by Merck Group (1 mentions)

Spelling variants of this product

EnzChek phosphatase assay kit (12 citations) Tyrosine (7 citations)

16 protocols using rediplate 96 enzchek tyrosine phosphatase assay kit

1

Quantifying PTPN6 Phosphatase Activity

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Phosphatase activity of PTPN6 was assessed using the RediPlate 96 EnzChek Tyrosine Phosphatase Assay kit (Molecular Probes, Eugene, OR, USA). After an overnight serum starvation, cells were washed with saline before lysis in buffer containing 50mM MOPS, 50mM NaCl, 1mM DTT and 0.3% Tween and protease inhibitor. Lysates were transferred into RediPlate wells and incubated for 30 min at 20-22° C before reading for fluorescence.
Demosthenous C., Han J.J., Hu G., Stenson M, & Gupta M. (2015). Loss of function mutations in PTPN6 promote STAT3 deregulation via JAK3 kinase in diffuse large B-cell lymphoma. Oncotarget, 6(42), 44703-44713.
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2

SHP-1 Phosphatase Activity Assay

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A RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (R-22067) was used for SHP-1 activity assay (Molecular Probes, Carlsbad, CA). The method was as described previously52 (link).
Su J.C., Mar A.C., Wu S.H., Tai W.T., Chu P.Y., Wu C.Y., Tseng L.M., Lee T.C., Chen K.F., Liu C.Y., Chiu H.C, & Shiau C.W. (2016). Disrupting VEGF-A paracrine and autocrine loops by targeting SHP-1 suppresses triple negative breast cancer metastasis. Scientific Reports, 6, 28888.
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3

Assessment of PTP1B Phosphatase Activity

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For in-vivo PTP1B phosphatase activity assessment, lysate of regorafenib-treated DLD1 cells was first incubated overnight with buffer containing with anti-PTP1B antibody for co-immunoprecipitation, In the next day, we added protein G-Sepharose 4 Fast Flow (GE Healthcare Bio-Science, NJ) into each sample and incubated for another 3 hours. The samples were kept at 4 °C condition with rotation. For ex-vivo assessment, recombinant PTP1B proteins (Genway, GWB-IE1774) was first treated with regorafenib for 4 hours and assessed by a commercial kit, RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (R-22067) for PTP1B activity assay (Molecular Probes, Invitrogen, CA).
Teng H.W., Hung M.H., Chen L.J., Chang M.J., Hsieh F.S., Tsai M.H., Huang J.W., Lin C.L., Tseng H.W., Kuo Z.K., Jiang J.K., Yang S.H., Shiau C.W, & Chen K.F. (2016). Protein tyrosine phosphatase 1B targets PITX1/p120RasGAP thus showing therapeutic potential in colorectal carcinoma. Scientific Reports, 6, 35308.
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4

Sorafenib, Brivanib, and Sunitinib Effects on SHP-1 Activity

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After treatment with sorafenib, brivanib, or sunitinib, HCC protein extract was incubated with anti-SHP-1 antibody in immunoprecipitation buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1% NP-40, and 1% sodium deoxycholate) overnight. Protein G-Sepharose 4 Fast flow (GE Healthcare Bio-Science, NJ) was added to each sample, followed by incubation for 3 h at 4°C with rotation. A RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (R-22067) was used for SHP-1 activity assay (Molecular Probes, Invitrogen, CA).
Tai W.T., Hung M.H., Chu P.Y., Chen Y.L., Chen L.J., Tsai M.H., Chen M.H., Shiau C.W., Boo Y.P, & Chen K.F. (2016). SH2 domain-containing phosphatase 1 regulates pyruvate kinase M2 in hepatocellular carcinoma. Oncotarget, 7(16), 22193-22205.
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5

Quantifying SHP-1 Phosphatase Activity

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A RediPlate 96 EnzChek® Tyrosine Phosphatase Assay Kit (R-22067) was used for SHP-1 activity assay (Molecular Probes, Invitrogen, CA). To detect the phosphatase activity of endogenous SHP-1, SHP-1 protein in Huh-7 cell lysates was incubated with anti-SHP-1 antibody at 4 °C overnight and precipitated with Protein G-Agarose beads (Roche). The beads were washed with immunoprecipitation buffer (20 mM Tris-HCl (pH 8.0), 50 mM NaCl, 0.5% Triton-X-100, and 10% glycerol) and placed into RediPlate wells. SHP-1 activity was measured at 360/40 and 460/40 nm. The amount of SHP-1 protein used for the phosphatase assay were evaluated by western blotting. For in vitro phosphatase assay, 4 nM recombinant His-SHP-1 or His-SHP-1Δ517–597 protein and 4 nM HNF1A-AS1 were added into RediPlate wells and incubated for 30 min at RT before reading fluorescence.
Ding C.H., Yin C., Chen S.J., Wen L.Z., Ding K., Lei S.J., Liu J.P., Wang J., Chen K.X., Jiang H.L., Zhang X., Luo C, & Xie W.F. (2018). The HNF1α-regulated lncRNA HNF1A-AS1 reverses the malignancy of hepatocellular carcinoma by enhancing the phosphatase activity of SHP-1. Molecular Cancer, 17, 63.
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6

SHP-1 Phosphatase Activity Assay

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The Rediplate 96 EnzChek®Tyrosine Phosphatase Assay Kit (R-22067) was used for SHP-1 activity assay (Molecular Probes, Carlsbad, CA). The SHP-1 phosphatase activity was measured as described before [30 (link)].
Kong J., Kong F., Gao J., Zhang Q., Dong S., Gu F., Ke S., Pan B., Shen Q., Sun H., Zheng L, & Sun W. (2014). YC-1 enhances the anti-tumor activity of sorafenib through inhibition of signal transducer and activator of transcription 3 (STAT3) in hepatocellular carcinoma. Molecular Cancer, 13, 7.
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7

SHP-1 Tyrosine Phosphatase Assay

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SHP-1 was immunoprecipitated with 2 μg anti-SHP-1 polyclonal antibody (Santa Cruz) from control and Shp-1-Tg VSMCs overnight, 20 μl Protein A/G PLUS-Agarose (Santa Cruz) was added and beads were washed and then assayed by RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (Molecular Probes, Eugene, OR, USA).
Qi W., Li Q., Liew C.W., Rask-Madsen C., Lockhart S.M., Rasmussen L.M., Xia Y., Wang X., Khamaisi M., Croce K, & King G.L. (2016). SHP-1 activation inhibits vascular smooth muscle cell proliferation and intimal hyperplasia in a rodent model of insulin resistance and diabetes. Diabetologia, 60(3), 585-596.
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8

Knockdown of SHP-1 Phosphatase Activity

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siRNAs targeting SHP-1 and scrambled control siRNAs were purchased from Dharmacon (Thermo Scientific, Chicago, IL, USA). Cells were transfected with siRNAs using Lipofectamine 2000 (Invitrogen Life Technologies, Rockville, MD, USA) to knock down gene expression. For SHP-1 activity assay, a RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (Molecular Probes, Carlsbad, CA, USA) was used following the manufacturer’s instructions.
Liu C.Y., Huang T.T., Chu P.Y., Huang C.T., Lee C.H., Wang W.L., Lau K.Y., Tsai W.C., Chao T.I., Su J.C., Chen M.H., Shiau C.W., Tseng L.M, & Chen K.F. (2017). The tyrosine kinase inhibitor nintedanib activates SHP-1 and induces apoptosis in triple-negative breast cancer cells. Experimental & Molecular Medicine, 49(8), e366-.
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9

SHP-1 Phosphatase Activity Assay

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A RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (R-22067) was used for the SHP-1 activity assay (Molecular Probes, Carlsbad, CA). The method was as described previously [16] (link).
Su J.C., Tseng P.H., Wu S.H., Hsu C.Y., Tai W.T., Li Y.S., Chen I.T., Liu C.Y., Chen K.F, & Shiau C.W. (2014). SC-2001 Overcomes STAT3-mediated Sorafenib Resistance through RFX-1/SHP-1 Activation in Hepatocellular Carcinoma. Neoplasia (New York, N.Y.), 16(7), 595-605.
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10

SHP-1 Phosphatase Activity Assay

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A RediPlate 96 EnzChek Tyrosine Phosphatase Assay Kit (Molecular probe, R-22067) was used for SHP-1 activity assay. The method was as described previously [43 (link)].
Su J.C., Tseng P.H., Hsu C.Y., Tai W.T., Huang J.W., Ko C.H., Lin M.W., Liu C.Y., Chen K.F, & Shiau C.W. (2014). RFX1–dependent activation of SHP-1 induces autophagy by a novel obatoclax derivative in hepatocellular carcinoma cells. Oncotarget, 5(13), 4909-4919.
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