Cell Signaling Technology

Product Pathways - Protein Stability

PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Kit #5562

motif   ub   ubiscan  


No. Size Price
5562S 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® Ubiquitin Branch Motif (K-ε-GG) 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). UbiScan® Technology developed at Cell Signaling Technology is a unique strategy for global analysis of cellular ubiquitination. In this assay, the PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Antibody Bead Conjugate, a proprietary ubiquitin branch (“K-ε-GG”) antibody with specificity for a di-glycine tag that is the remnant of ubiquitin left on protein substrates after trypsin digestion, is used to enrich ubiquitinated peptides from trypsin digested cell samples. This enrichment is followed by LC-MS/MS analysis for quantitative profiles of hundreds to over a thousand non-redundant ubiquitinated sequences. More specifically, cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and resulting peptides are fractionated by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using Ubiquitin Remnant Motif (K-ε-GG) Antibody coupled to protein A agarose beads. Unbound peptides are removed through washing, and K-GG-motif-containing peptides are eluted with dilute acid. Reversed-phase chromatography is performed on microtips to separate phospho-peptides 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。UbiScan®技术由CST公司开发,是全面分析细胞泛素化过程的独特策略。在此分析中,PTMScan® Ubiquitin Remnant Motif (K-ε-GG) Antibody Bead Conjugate作为一个泛素分支(“K-ε-GG”)抗体,对胰蛋白酶消化后泛素留在蛋白底物上的二甘氨酸标签具有特异性,可以用来富集胰蛋白酶消化的细胞样本中的泛素化肽段。富集后经LC-MS/MS分析确认可识别数百到上千个非冗余的泛素化序列。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。肽段然后使用Ubiquitin Remnant Motif (K-ε-GG) Antibody结合到蛋白A琼脂糖珠子上进行免疫亲和纯化。未结合的抗体被清洗时去除,包含K-GG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。

Motif Logo

Motif Logo

Motif analysis was done using all ubiquitin remnant peptides in a ubiquitin remnant motif antibody PTMScan® study. Cellular tryptic peptides from HCT116 cells. untreated or treated with MG132, were immunoprecipitated with ubiquitin remnant motif antibody K-ε-GG and analyzed by OrbiTrap MS. The study gave 599 non-redundant sites. The motif logo shows that the K-ε-GG antibody is a general motif antibody that recognizes the K-ε-GG motif and does not have other amino acid preferences.

使用全部泛素残余抗体进行的PTMScan研究中发现的泛素残余肽段进行模体分析。HCT116细胞的细胞胰蛋白酶肽,未处理或MG132处理,与泛素残余模体抗体K-ε-GG进行免疫共沉淀,然后通过OrbiTrap MS分析。此研究给出599个非冗余位点。模体标识图显示K-ε-GG抗体是一个识别K-ε-GG模体的全模体抗体,对其它氨基酸没有偏好。

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。


PTMScan® technology employs a proprietary methodology from Cell Signaling Technology 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®). UbiScan® is a powerful method utilizing a unique ubiquitin-branch antibody for ubiquitinated sequence identification and comprehensive analysis of cellular ubiquitin pathways. Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (2-4). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25a and bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (5-8).


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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