Cell Signaling Technology

Product Pathways - Screening Technologies

PTMScan® Phospho-ST*P Motif (ST*P) XP® Rabbit mAb Kit #5566

motif   phospho-threonine   threonine  


No. Size Price
5566S 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-PLK Binding Motif (ST*P) Immunoaffinity Beads 80 µl Rabbit
PTMScan® IAP Buffer (10X) #9993 600 µl
PTMScan® Limited Use License license


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® (ST*P) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing the ST*P motif (T* = phospho-threonine). 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-ST*P Motif (ST*P) XP® Rabbit mAb coupled to protein A 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. Note: The XP® monoclonal antibody in this kit was generated using our proprietary XMT® monoclonal technology.

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®(ST*P) Motif Antibody珠子可以用来特异性富集磷酸化的ST*P基序(T*是磷酸化苏氨酸)。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-ST*P Motif (ST*P) XP® Rabbit mAb结合到蛋白A琼脂糖珠上免疫亲和纯化。未结合的抗体被清洗时去除,包含磷酸化模体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。注意:此试剂盒中的XP®单克隆抗体利用我们所有的XMT®单克隆技术开发。



Chart showing the proportion of underlying sequence motifs found in a PTMScan® study using a PLK binding motif antibody and OrbiTrap MS analysis. Analysis of peptides from HeLa cells, untreated and nocodazole-treated, gave 452 non-redundant sites containing a PLK binding and related motifs. The primary motif is highlighted in white 98% of these sites fit the ST*P motif; only 2% of the sites are from other phospho-Ser/Thr containing peptides that do not have the motif, which shows that this antibody is very specific for the ST*P motif.

图片显示使用PLK结合模体抗体和Orbitrap MS分析进行的PTMScan研究中发现的序列模体的属性. 对未处理或nocodazole处理的HeLa细胞肽段的分析给出包含PLK结合模体和相关模体的452个非冗余位点。主要的模体用白色高亮标出。98%的位点符合ST*P模体;只有2%的文电来自其它包含磷酸化丝氨酸/苏氨酸而非此模体的肽段,这表明此抗体对ST*P模体有很高特异性。

Motif Logo

Motif Logo

The Motif Logo shows the amino acid distributions around the sites recognized by the antibody.


Western Blotting

Western Blotting

Plk bing StP motif western blot analysis of extracts from HeLa cells, untreated or Nocodazole treated, using Phospho-ST*P Motif (ST*P) XP® Rabbit mAb.

使用Phospho-ST*P Motif (ST*P) XP® Rabbit mAb对来自未处理或Nocodazole处理的Hela细胞抽提液进行Plk bing StP模体的Western Blot分析。

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。


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 phosphopeptides, 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). The ST*P motif is implicated in signaling by polo-like kinases (PLKs). These Ser/Thr protein kinases play essential roles during the cell cycle. At least four PLKs exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4 (2). PLKs have a highly conserved N-terminal kinase domain and a relatively divergent C-terminal domain called the Polo-box domain (PBD). Of the four PLKs, PLK1 is the best characterized (3). PLK1 functions as a key regulator of mitotic events by phosphorylating substrate proteins on centrosomes, kinetochores, the mitotic spindle, and the midbody, and is crucial for proper progression through multiple stages of mitosis (4-6). The PBDs of PLK1 function as a phospho-Ser/Thr-binding module, optimally recognizing the sequence motif S[S*/T*][P/X] (PLK1 binding motif) (6). Binding of phosphopeptides containing S[S*/T*][P/X] motif by the PBD in PLK1 relieves its inhibitory function on kinase activity (7). S[S*/T*]P peptides are phosphorylated by proline-directed kinases such as cyclin dependent kinases (CDKs) (6). These findings imply that priming phosphorylations on substrates or docking proteins by other mitotic kinases such as CDKs may target PLK1 to its substrates and simultaneously activate its kinase domain. The ST*P Motif (ST*P) XP® Rabbit mAb contained in this kit, which binds to ST*P, was developed by Cell Signaling Technology and is a useful tool to study PLK1 binding proteins and PLK1 substrates.

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)基序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)。ST*P基序在polo样激酶(PLK)信号通路中提及。这些丝氨酸/苏氨酸蛋白激酶在细胞周期中扮演重要角色。哺乳动物细胞中至少存在四种PLK:PLK1、PLK2、PLK3和PLK4(2)。PLK具有高度保守的氨基末端激酶结构域和一个相对不保守的羧基末端结构域Polo-box域(PBD)。在四种PLK中,PLK1是描述得最清楚的(3)。PLK1作为有丝分裂事件的关键调节者而发挥作用,它可以磷酸化中心体、着丝粒、有丝分裂纺锤体、中心颗粒体的底物蛋白,对有丝分裂多个阶段的正确进展起关键作用(4-6)。PLK1的PBD作为磷酸化丝氨酸/苏氨酸结合基序而发挥作用,识别最优识别基序S[S*/T*][P/X](PLK1结合基序)(6)。PLK1的PBD结合包含S[S*/T*][P/X]模体的磷酸化肽段后解除其对激酶活性抑制功能(7)。S[S*/T*]P肽被脯氨酸导向的激酶如细胞周期依赖激酶(CDK)磷酸化(6)。这些发现提示其它有丝分裂激酶如CDK对底物或接合蛋白可能是通过PLK1和其底物作用而同时激活其激酶活性而促发的。包含在试剂盒中的ST*P Motif (ST*P) XP® Rabbit mAb可以结合ST*P,由CST公司开发,是研究PLK1结合蛋白和PLK1底物的有用工具。

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!

Companion Products

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.

XP is a registered 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.

用户评论 --- 共 0


我要参与评论 :