Specifications

“My lab has successfully used the agarose-TUBEs. LifeSensors’ TUBEs are easy to use and highly specific. My lab is looking forward to using the linkage specific TUBEs.”

Prof. David Wallach, The Weizmann Institute of Science

Based on protein domains known to possess an affinity for ubiquitin, Tandem Ubiquitin Binding Entities (TUBEs) have been developed for the isolation and identification of ubiquitylated proteins. TUBEs display up to a 1000-fold increase in affinity for polyubiquitin moieties over the single ubiquitin binding associated domain (UBA). In addition, TUBEs protect polyubiquitinated proteins from deubiquitination and proteasomal degradation, allowing for detection at relatively low abundance. These properties effectively allow TUBEs to “capture” protein in its polyubiqutinated state. TUBEs can be used to isolate, enrich, and identify ubiquitinated proteins from cell and tissue extracts.

Polyubiquitin chains are labile, particularly in cell lysates. Success with Agarose TUBEs is often dependent on the inclusion of deubiquitylase (DUB) inhibitors in your lysis buffers to prevent the removal of polyubiquitin chains and subsequent loss of polyubiquitylated proteins during your purification. LifeSensors offers PR619 (Cat. # SI9619), a non-selective inhibitor of cysteine-type DUBs, and 1,10-phenanthroline (Cat. # SI9649), a Zn2+ chelator and potent inhibitor of JAMM-type DUBs, for inclusion in your lysis buffer and protection of polyubiquitinated proteins during purification. Please see the agarose-TUBE manual for further details.

TUBE 1 is based on UBAs from the protein ubiquilin. Agarose coupled TUBEs allow for efficient recovery of polyubiquitinated proteins in a single step. Polyubiquitinated protein can easily be separated from TUBEs for proteomic studies or identification by Western blotting.

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Info

Size	1 ml of a 50% slurry
Buffer	PBS, pH 7.2, 20% ethanol
Recommended Concentration	10-20 µL of resin in 500 µL lysate (see manual)
Physical State	Agarose-TUBE 1 is provided as a 50% slurry
Storage	-20°C. Storage below this temperature may result in freezing and/or loss of function.

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。