Cell Signaling Technology

Product Pathways - Screening Technologies

PTMScan® Phospho-MAPK/CDK Substrate Motif (PXS*P and S*PXK/R) Kit #4652



No. Size Price
4652S 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-MAPK/CDK Substrate Motif (PXS*P, S*PXK/R) 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® (PXS*P or S*PXK/R) Motif Antibody bead conjugates are used to specifically enrich phosphopeptides containing the PXS*P motif or the S*PXK/R motif (S* = phospho-serine). 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-MAPK/CDK Substrates (PXSP or SPXR/K) Rabbit mAb #2325 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.

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。这种方法,PTMScan® (PXS*P or S*PXK/R) Motif Antibody基序抗体珠可以用来特异性富集磷酸化的PXS*P模体或S*PXK/R模体(S*是磷酸化丝氨酸)。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-MAPK/CDK Substrates (PXSP or SPXR/K) Rabbit mAb #2325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。一个有限使用许可证批准了使用这个试剂盒中的受专利保护的PTMScan方法。



Chart showing the proportions of underlying sequence motifs found in a PTMScan® study using a MAPK/CDK substrate motif antibody and OrbiTrap MS analysis. Analysis of peptides from HeLa cells, untreated (experiment #10684) and nocodazole-treated (experiment #10685), gave 439 nonredundant sites containing MAPK/CDK substrate and related motifs. 20% of these sites are motif PXS*PX(R/K); 70% are motif PXS*P; and 10% are motif S*PX(R/K). The antibody is phospho-serine specific and does not recognize phospho-threonine peptides.

图片显示使用MAPK/CDK底物模体抗体和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(experiment #10684)或nocodazole处理(experiment #10685)的HeLa细胞肽段的分析给出包含MAPK/CDK底物模体和相关模体的439个非冗余位点。这些位点中20%为PXS*PX(R/K),70%为PXS*P模体,10%为S*PX(R/K)模体。此抗体是磷酸化特异性的,与包含磷酸化苏氨酸的肽段或蛋白没有交叉反应。

Motif Logo

Motif Logo

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


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® is 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 MAPK and CDK families of serine/threonine protein kinases play important roles in proliferation and cell cycle control. These kinases phosphorylate threonine or serine followed by a proline residue (2-4). MAPK phosphorylates substrates with the consensus sequence PX(S/T)P, and CDKs phosphorylate substrates containing the consensus sequence (S/T)PXK/R. Phospho-MAPK/CDK Substrates (PXSP or SPXR/K) Rabbit mAb #2325, contained in this kit, was developed at CST for use in the study and discovery of new MAPK and CDK substrates (5,6).

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分析磷酸化水平的改变(1)。MAPK和CDK丝氨酸苏氨酸蛋白激酶家族在细胞增殖和周期调控中扮演重要角色。这些激酶磷酸化跟随有脯氨酸的苏氨酸或丝氨酸(2-4)。MAPK磷酸化含有一致序列PX(S/T)P的底物,CDK磷酸化含有一致序列(S/T)PXK/R的底物。包含在试剂盒中的Phospho-MAPK/CDK Substrates (PXSP or SPXR/K) Rabbit mAb #2325,由CST公司开发,用以研究新的MAPK和CDK激酶底物(5,6)。

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.

U.S. Patent No. 5,675,063.

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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