Specifications

This kit allows you to detect whether your target protein is modified by K63 poly-ubiquitin.

Determining the linkage of polyubiquitin on target proteins is challenging. The traditional methods are either through Mass Spectrometry or immunoblot with linkage specific antibodies, which are cumbersome. LifeSensors has developed a more definitive method for demonstrating the ubiquitylation linkage of a protein, which is to couple immunoprecipitation of polyubiquitylated protein with digestion by a linkage specific deubiquitylase prior to immunoblot analysis. An increased signal for the unmodified substrate or a decreased signal of polyubiquitylated substrate at high molecular weight after K48/K63 specific DUB treatment is a clear indication that the protein was K48/K63 ubiquitylated. We have built this kit around the use of Tandem Ubiquitin Binding Entities (TUBEs) which bind to all ubiquitin chain linkages.

This kit includes the following DUBs:

• A Lys63 linkage specific DUB – 35 µg, 10 µM

• A broad spectrum DUB – 25 µg, 10 µM

This product is protected by one or more US or Foreign patents. Please read the Limited Use Label License to learn more. By purchasing this product, the purchaser agrees to comply with these terms.

Info

Kit Contents:

Tube 1	Anti-Ub-TUBE1 or TUBE2, Agarose – 1mL of a 50% slurryBuffer: PBS, pH 7.2, 20% ethanolStorage: -20°C. Avoid storage at lower temperature. Small batches of equilibrated resin can be stored at 4°C for up to 1 week
Tube 2	Elution wash buffer – 2mL
Tube 3	Elution buffer – 1mL
Tube 4	Neutralization buffer (10X) – 0.5mL
Tube 5	USP2core – 25ugStorage: -80°C

Life Sensors的泛素和泛素链泛素是一种小的多肽，通过其C端与目标蛋白上的赖氨酸的ε-氨基缀合。这种缀合称为单泛素化。随后，可以利用遍在蛋白表面上的七个赖氨酸残基（K6，K11，K27，K29，K33，K48，K63）中的任何一个，将额外的遍在蛋白部分与初始遍在蛋白结合。这种泛素链形成称为多泛素化。泛素的酶结合是通过一系列酶进行的，即泛素激活酶E1，泛素结合酶E2和泛素连接酶E3。蛋白质的泛素化在细胞中是可逆的。单泛素化和多泛素化的链均被去泛素化酶催化的水解作用裂解 （DUB）。LifeSensors在遍在泛素途径的所有水平的酶（包括E1，E2和E3酶和DUB）上都有丰富的经验。LifeSensors对蛋白质纯化的广泛了解使我们能够生产大约30种E2酶，20种E3酶和35个DUB，并具有定制表达的能力。此外，我们广泛的检测方法简化了确认酶活性和确定E2-E3对兼容性的过程，并确保您可以选择最能解决希望探索的特定，关键生物学问题的酶。LifeSensors已为您的研究开发了多种泛素衍生物，包括用于结合相互作用研究的DUB耐药性聚泛素链，用于DUB分析和筛选的泛素结合物（荧光团和报告酶），选择性泛素链（二，三和四环素）泛素链）用于DUB特异性和结合测定，位点特异性赖氨酸泛素突变体用于E3连接酶和DUB选择性测定。