Cell Signaling Technology

Product Pathways - Motif Antibodies

PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Kit #12170

CK2   Motif Antibody   PhosphoScan   PTMScan  


No. Size Price
12170S 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-CK2 Substrate Motif (S*/T*DXE) Immunoaffinity Beads 80 µl Rabbit
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。这些蛋白质翻译后修饰包括磷酸化(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

The Motif Logo was generated from a PhosphoScan® LC-MS/MS experiment using 471 nonredundant tryptic peptides derived from Jurkat cells treated with Calyculin A #9902 and pervanadate, and immunoprecipitated using PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Immunoaffinity Beads. This PhosphoSitePlus® logo represents the enrichment of an amino acid at a residue position relative to the abundance of that amino acid in the background, in this case, the human proteome. Residues represented at the top of the logo are the most enriched, whereas those below the y-axis occur less frequently in the sample than would be expected in the proteome. The PSP logo reflects the specificity of the antibody for the S*/T*DXE motif. 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Calyculin A #9902 及过钒酸钠处理的Jurkat细胞的471 低丰度胰蛋白酶消化的多肽,用PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Immunoaffinity Beads进行免疫沉淀。PhosphoSitePlus®标志代表一个氨基酸在一个残基位置上的相对于背景氨基酸丰度的富集,在该图中背景为人类蛋白质组。在logo上方的残基代表最丰富的,而那些y轴以下的表示比其在蛋白质组中的预期还要更少出现。PSP logo反应了S*/T*DXE motif抗体的特异性。 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 underlying CK2 substrate-like motifs in a PhosphoScan® LC-MS/MS experiment of tryptic peptides generated from Jurkat cells treated with Calyculin A #9902 and pervanadate, and immunoprecipitated using PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Immunoaffinity Beads. Of the 354 motif-containing peptides, 54% contained phospho-serine and 46% contained phospho-threonine. The proportion of peptides with aspartate or glutamate at the +1 position is 83% and at the +3 position is 73%. Peptides with aspartate or glutamate at both the +1 and +3 positions is 63%.

图片显示了从PhosphoScan® LC-MS/MS实验中来源于Calyculin A #9902 及过钒酸钠处理的Jurkat细胞,并被胰蛋白酶消化产生的多肽中的CK2 substrate-like motif。免疫沉淀中使用的是PTMScan® Phospho-CK2 Substrate Motif (S*/T*DXE) Immunoaffinity Beads。在这些含有354个motif的多肽中,54%含有磷酸化丝氨酸,46%含有磷酸化Thr。多肽中含有Asp或Glu的比例是+1位置83%,+3位置73%。+1和+3都有包含Asp和Glu的多肽比例是63%。

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). Casein Kinase II (CK2) is a highly conserved, ubiquitously expressed, and constitutively active tetrameric serine/threonine protein kinase with hundreds of substrates participating in the regulation of a variety of cellular processes including cell cycle progression, apoptosis, transcription, inflammation, and the DNA damage response. Research studies have implicated CK2 in roles related to viral infection, cancer and other diseases (2-6). CK2 substrates contain multiple acidic residues (Asp and Glu) located downstream of the phosphorylated serine or threonine residue. The consensus sequence for CK2 substrates is pS/pTD/EXD/E with the most crucial residue at the +3 position, followed by the residue at the +1 position (7).PhosphoScan®采用磷酸化残基(Tyr、Ser、Thr)motif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)。Casein Kinase II (CK2)是一个高度保守、广泛表达,具有组成型活性的四聚体Ser/Thr 蛋白激酶,具有多种参与细胞周期进程、凋亡、转录、炎症以及DNA损伤应激等一系列细胞进程调控的底物。研究发现,CK2与病毒感染、癌症及其他疾病都相关(2-6)。CK2底物包含多种酸性残基(Asp和 Glu),位于磷酸化Ser或Thr残基的下游。CK2底物的共有序列是pS/pTD/EXD/E,最重要的残基在+3 位点,随后是+1 位点的残基(7)。

Application References

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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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