PTMScan ® Phospho-Tyrosine Rabbit mAb (P-Tyr-1000) Kit

品牌： CST

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产品信息

简单描述

PTMScan for studying Tyrosine phosphate in the research area.

商品描述

Product Usage Information

Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit.

背景

PhosphoScan® Technology employs phospho-residue (Tyr, Ser, Thr) motif antibodies for phosphopeptide immunoaffinity purification from cell extracts combined with LC tandem MS/MS to identify and quantify changes in phosphorylation levels (1). Tyrosine phosphorylation plays a key role in cellular signaling. Research studies have shown that in cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (2). Antibodies specific for phospho-tyrosine have been invaluable reagents in these studies (3,4). The Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954, developed by Cell Signaling Technology, provides an exceptionally sensitive tool for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery. PTMScan® (Y 1.Rush, J. et al. (2005) Nat Biotechnol 23, 94-101. 2.Schlessinger, J. (2000) Cell 103, 211-25. 3.Blume-Jensen, P. and Hunter, T. (2001) Nature 411, 355-65. 4.Ward, S.G. et al. (1992) J Biol Chem 267, 23862-9.

制备和贮存

保存方式

Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20°C. Do not aliquot the antibody.

参考图片

Motif analysis was done using all tyrosine phosphopeptides generated from a PhosphoScan® experiment of mouse brain and mouse liver tryptic peptides using PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads. The Motif Logo, drawn from 1178 nonredundant sites, shows the amino acid distributions around the sites recognized by Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954. The motif logo shows that this antibody is a general phospho-tyrosine antibody and does not have amino acid preferences at other positions.

利用所有的酪氨酸磷酸化多肽进行Motif分析，多肽来自于PhosphoScan®实验中使用PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads沉淀到的小鼠大脑和小鼠肺胰蛋白酶消化的多肽。这个Motif Logo，从1178个低丰度位点中获得，显示了被Phospho-Tyrosine (P-Tyr-1000) Rabbit mAb #8954识别位点周围的氨基酸分布。该Logo显示，该抗体是一个总的磷酸化酪氨酸抗体，对于其他位点没有氨基酸倾向性。

Representative motif analysis of 2362 phospho-tyrosine tryptic peptides generated from a PhosphoScan® experiment of Jurkat cells treated with Calyculin A #9902 and pervanadate using PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads. Motif logo was generated using the PhosphoSitePlus® Logo Generator and reflects relative frequency of residues in each position. For more information, please visit PhosphoSitePlus® at www.phosphosite.org.

有代表性的2362种酪氨酸磷酸化胰蛋白酶消化的多肽进行的Motif分析，这些多肽来自于PhosphoScan®实验中使用PTMScan® Phospho-Tyrosine Motif (Y*) (P-Tyr-1000) Immunoaffinity Beads沉淀到的JurKat细胞。Motif Logo使用PhosphoSitePlus® Logo Generator生成，反映了每个位置上的残基相对频率。更多信息，请访问PhosphoSitePlus® at www.phosphosite.org.

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8803S

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