Cell Signaling Technology

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PTMScan® Phospho-ATM/ATR Substrate Motif [pSQ] Kit #12267


No. Size Price
12267S 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-ATM/ATR Substrate Motif [pSQ] Immunoaffinity Beads 80 µl Rabbit IgG
PTMScan® IAP Buffer (10X) #9993 600 µl
PTMScan® Limited Use License license


PTMScan® Technology employs a proprietary methodology from Cell Signaling Technology for peptide enrichment by immunoprecipitation using a specific bead-conjugated antibody in conjunction with liquid chromatography (LC) tandem mass spectrometry (MS/MS) for quantitative profiling of post-translational modification (PTM) sites in cellular proteins. These include phosphorylation (PhosphoScan®), ubiquitination (UbiScan®), acetylation (AcetylScan®), and methylation (MethylScan®), among others. PTMScan® Technology enables researchers to isolate, identify, and quantitate large numbers of post-translationally modified cellular peptides with a high degree of specificity and sensitivity providing a global overview of PTMs in cell and tissue samples without preconceived biases about where these modified sites occur. For more information on PTMScan® Proteomics Services, please visit www.cellsignal.com/services/index.html.PTMScan®技术使用的是CST用于多肽富集的专利技术,利用免疫共沉淀通过一种特殊的与bead偶联的抗体结合液相色谱(LC)串联质谱(MS/MS)来定量分析细胞内蛋白质翻译后修饰(PTM)位点。此外还有磷酸化(PhosphoScan®)、泛素化(UbiScan®)、乙酰化 (AcetylScan®)以及甲基化(MethylScan®)。 PTMScan®技术可以使研究者能够分离、鉴定以及定量许多翻译后修饰的细胞内多肽,并具有很高的特异性和灵敏度,从而了解细胞和组织样品中PTM的全面概况,并且不带有任何关于这些修饰位点位于何处的先入为主的偏见。更多有关PTMScan® Proteomics Services的信息,请访问www.cellsignal.com/services/index.html.

Motif Logo

Motif Logo

This Motif Logo was generated from a PhosphoScan® LC-MS/MS experiment using 1111 nonredundant tryptic peptides derived from Jurkat cells treated with Calyculin A #9902 and pervanadate and immunoprecipitated using PTMScan® Phospho-ATM/ATR Substrate Motif [pSQ] Immunoaffinity Beads. The PhosphoSitePlus® logo reflects the relative prevalence of an amino acid in each position relative to the phospho-serine background in the human proteome. Residues represented above the x-axis are enriched relative to their expected frequency in this background. For more information on motif analysis using PSP, please visit www.phosphosite.org.

Motif Logo出自PhosphoScan® LC-MS/MS实验,实验中使用来源于经Calyculin A #9902 和 pervanadate处理的Jurkat细胞的1111种低丰度胰蛋白酶消化的多肽,这些多肽使用PTMScan® Phospho-ATM/ATR Substrate Motif [pSQ] Immunoaffinity Beads免疫沉淀得到。该PhosphoSitePlus® logo代表了相对于人蛋白质组中的磷酸化丝氨酸的每个位置上氨基酸的相对频率。相对于背景中的预期,那些x轴以上的残基被富集。更多关于使用PSP进行motif 分析的信息,请访问www.phosphosite.org。



This chart shows the uderlying motif distribution within 1111 nonredundant tryptic peptides derived from an LC-MS/MS experiment using Jurkat cells treated with Calyculin A #9902 and pervanadate and immunoprecipitated with PTMScan® Phospho-ATM/ATR Substrate Motif [pSQ] Immunoaffinity Beads. Within the same data set, phospho-serine in the central position constitutes 88% of the peptides, while phospho-threonine constitutes 12%. The most frequent submotif is [pSQG] with a 16% rate of occurrence relative to the total data set and 28% relative to the subset with the pSQ motif.

图片显示了PhosphoScan® LC-MS/MS实验中得到的motif分布,实验中使用来源于经Calyculin A #9902 和 pervanadate处理的Jurkat细胞的1111种低丰度胰蛋白酶消化的多肽。免疫沉淀中使用的是PTMScan® Phospho-ATM/ATR Substrate Motif [pSQ] Immunoaffinity Beads。在相同的数据组中,中心位置磷酸化的丝氨酸占88%,而磷酸化的苏氨酸占12%。最频繁的submotif 是[pSQG],出现的比例相对于整个数据组为16%,相对于含有pSQ motif的亚组为28%。

Directions for Use

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. An alternate PTMScan® IAP Buffer Plus Detergent #9992, which may reduce nonspecific interactions, is available separately. A detailed protocol and Limited Use License allowing the use of the patented PTMScan® method are included with the kit.使用含尿素的缓冲液裂解细胞,细胞内蛋白被蛋白酶消化,产生的多肽通过反相固相萃取纯化。这些肽段然后使用偶联到蛋白A琼脂糖珠的PTMScan® Motif Antibody进行免疫亲和纯化。未结合的多肽通过清洗去除,捕捉到的含有PTM的肽段使用稀释的酸进行洗脱。在浓缩富集到的多肽用于LC-MS/MS 分析前,需要在microtip上进行反相纯化以脱盐并将多肽从抗体中分离。CST推荐使用试剂盒中提供的PTMScan® IAP Buffer #9993。另外还可以使用PTMScan® IAP Buffer Plus Detergent #9992缓冲液,它可以减少非特异性反应,可以单独购买。有关PTMScan®专利方法的详细操作步骤以及使用许可限制可在试剂盒中查询。


PhosphoScan® Technology employs a phospho-residue (Tyr, Ser, Thr) motif antibody for phospho-peptide immunoaffinity purification from cell extracts combined with LC tandem MS/MS to identify and quantify changes in phosphorylation levels (1). Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are related kinases that regulate cell cycle checkpoints and DNA repair (2). The identified substrates for ATM are p53, p95/NBS1, MDM2, Chk2, BRCA1, CtIP, 4E-BP1, and Chk1 (2,3). The consensus sequence for ATM/ATR substrates is [pS/pTQ]. Hydrophobic amino acids at positions -3 and -1, and negatively charged amino acids at position +1 are positive determinants for substrate recognition by these kinases. Positively charged residues surrounding the [pS/pTQ] are negative determinants for substrate phosphorylation (4). The complex phenotype of AT cells suggests that it likely has additional substrates (4). To better understand the kinase and identify substrates for ATM and the related kinase ATR, we have developed antibodies that recognize phosphorylated serine or threonine in the [pS/pTQ] motif.PhosphoScan® 技术采用磷酸化残基(Tyr、Ser、Thr)基序抗体对细胞抽提物进行磷酸化肽段免疫亲和纯化,结合液相色谱(LC)串联质谱(MS/MS)来鉴定和定量分析磷酸化水平的改变(1)。共济失调毛细血管扩张症突变蛋白激酶(ATM)和共济失调毛细血管扩张症Rad3相关激酶(ATR)是相关的激酶,与细胞周期检查点和DNA修复有关(1)。已识别的ATM底物包括p53、p95/NBS1、MDM2、Chk2、BRCA1、CtIP、4E-BP1和Chk1(2,3)。ATM/ATR底物的共有序列是[pS/pTQ]。-3和-1位的疏水氨基酸和+1位的负电荷氨基酸对这些激酶底物识别有正向决定作用。[pS/pTQ]附近的正电荷残基对底物磷酸化有负向作用(4)。AT细胞的复杂表型提示它们还有更多底物(4)。为了更好的理解激酶和识别ATM和ATR的相关底物,CST开发了识别[pS/pTQ]基序中磷酸化丝氨酸和苏氨酸的抗体。

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.

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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