Cell Signaling Technology

Product Pathways - Screening Technologies

PTMScan® Phospho-Tyrosine Mouse mAb (P-Tyr-100) Kit #5636

motif   py  

REACTIVITY

No. Size Price
5636S 1 Kit ( 10 assays ) 请询价 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
PTMScan® Phospho-Tyr Motif (Y*) Immunoaffinity Beads 80 µl Mouse
PTMScan® IAP Buffer (10X) #9993 600 µl
PTMScan® Limited Use License license

Description

PTMScan® technology allows for the identification of post-translational modification sites in cellular proteins by utilizing immunoprecipitation in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS). In this assay, PTMScan® (Y*) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing *Y motif (Y* = phospho-tyrosine). More specifically, cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and resulting peptides fractionated by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using Phospho-Tyrosine Mouse mAb (P-Tyr-100) # 9411 coupled to protein G agarose beads. Unbound peptides are removed through washing, and phospho-motif-containing peptides are eluted with dilute acid. Reversed-phase extraction is performed on microtips to separate phosphopeptides from antibody and to concentrate them for LC tandem MS. A Limited Use License allowing the use of the patented PTMScan method is provided with the kit.

PTMScan®技术利用免疫共沉淀结合液相色谱串联质谱来识别细胞内蛋白质的转录后修饰位点。在这个测试中PTMScan®(Y*) Motif Antibody珠子可以用来特异性富集磷酸化的*Y模体(Y*是磷酸化酪氨酸)。更特异地,细胞在含尿素的缓冲液中裂解,细胞内蛋白被蛋白被消化,产生被反相固相萃取的肽段碎片。这些肽段然后被Phospho-Tyrosine Mouse mAb (P-Tyr-100) # 9411结合到G蛋白琼脂糖珠上免疫亲和纯化。未结合的抗体被清洗时去除,包含磷酸化基序的肽段被稀释的酸洗脱。微探尖上进行反相固相萃取分离磷酸化肽段和抗体,然后浓缩以便进行液相色谱串联质谱。一个有限使用许可批准了使用这个试剂盒中的受专利保护的PTMScan方法。

Motif Logo

Motif Logo

Motif analysis was done using all tyrosine phosphopeptides generated from studies in the published report by Rikova, K.A., et al. (2007) Cell 131, 1190-1203. The Motif Logo, drawn from 4160 non-redundant sites, shows the amino acid distributions around the sites recognized by Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411. The motif analysis shows that this antibody is a general phospho-tyrosine antibody and does not have amino acid preferences at other positions.

使用Rikova, K.A.等发表的研究中产生的所有酪氨酸磷酸化肽段进行的模体分析。[Rikova, K.A., et al.(2007) Cell 131, 1190-1203]从4160个非冗余的位点画出的模体标志图显示出分布在Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411识别位点周围的氨基酸。模体分析显示此抗体是一个通用酪氨酸抗体,对其它位置的氨基酸没有偏好。

Directions for Use

The use of PTMScan® IAP Buffer Plus Detergent is recommended; it may reduce non-specific interactions. If detergent interferes with analyses to be performed on the IAP supernatant, use PTMScan® IAP Buffer. Please see the protocol in the kit package for further details.

推荐使用PTMScan® IAP Buffer Plus Detergent,它可以减少非特异性反应。如果去垢剂干扰IAP®上清液的分析,可使用PTMScan® IAP缓冲液。更多信息请参看工具和包装中的方案手册。

Background

PTMScan® technology employs a proprietary methodology from Cell Signaling Technology (CST) for antibody-based peptide enrichment combined with tandem mass spectrometry for quantitative profiling of post-translational modifications (PTMs), including phosphorylation (PhosphoScan®), ubiquitination (UbiScan®) and acetylation (AcetylScan®). PhosphoScan enables researchers to isolate a large number of cellular phospho-peptides, providing a global overview of phosphorylation in cell and tissue samples. This technology allows for the exploration of phosphorylation events, without preconceived biases about where phosphorylation sites will be found. PhosphoScan employs phospho-residue (Tyr, Ser, Thr) motif antibodies for phosphopeptide immunoaffinity purification from cell extracts combined with LC tandem MS to identify and quantify changes in phosphorylation levels (1). Tyrosine phosphorylation plays a key role in cellular signaling (2). In cancer, unregulated tyrosine kinase activity can drive malignancy and tumor formation by generating inappropriate proliferation and survival signals (3). Antibodies specific for phospho-tyrosine (4,5) have been invaluable reagents in these studies. The Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411, developed by Cell Signaling Technology, provides an exceptionally sensitive tools for studying tyrosine phosphorylation and monitoring tyrosine kinase activity in high throughput drug discovery.

PTMScan®技术采用CST公司独有的基于抗体的肽段富集方法,结合串联质谱定量分析翻译后修饰(PTM),包括磷酸化(PhosphoScan®)、泛素化(UbiScan®)和乙酰化(AcetylScan®)。PhosphoScan可以是研究者能够分离大量细胞内磷酸化肽,对细胞和组织的磷酸化情况有全局的认识。此技术使得我们可以探索磷酸化事件,不带有关于磷酸化位点在何处的先入为主的偏见。PhosphoScan采用磷酸化残基(Tyr、Ser、Thr)模体抗体对细胞抽提物进行磷酸化肽免疫亲和纯化,结合液相色谱串联质谱识别定量描述磷酸化水平的改变(1)。赖氨酸磷酸化在细胞信号转导中扮演重要角色(2)。在肿瘤中,上调酪氨酸激酶活性可以通过产生不适当的增生和存活信号而导致恶性肿瘤和肿瘤形成(3)。具有磷酸化酪氨酸特异性的抗体在这些研究中很有价值(4,5)。Cell Signaling Technology开发的Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411在高通量药物发现中检测酪氨酸磷酸化和酪氨酸激酶活性研究中是非常灵敏的新工具。

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

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For Research Use Only. Not For Use In Diagnostic Procedures.

Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PTMScan is a trademark of Cell Signaling Technology, Inc.

AcetylScan is a trademark of Cell Signaling Technology, Inc.

UbiScan is a trademark of Cell Signaling Technology, Inc.

MethylScan is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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