Specifications

IQF DiUb TechnologyLifeSensors’ diubiquitin substrates represent a new class of substrates for the continuous fluorescent measurement of true isopeptidase activity. The C-terminus of wild type ubiquitin is conjugated via an isopeptide bond to lysine 48 (K48) of a second ubiquitin molecule with the resultant diubiquitin forming an internally quenched fluorescent FRET pair (IQF). Each ubiquitin is labeled with a single molecule of either a fluorescent reporter (i.e. TAMRA) or a highly efficient quenching dye. Cleavage of the IQF DiUb by deubiquitylases leads to separation of the fluorophore from quencher and subsequent increase in observed fluorescence.

IQF DiUb VariantsThere is a remarkable difference between K48- and K63-linked ubiquitin molecules in their three-dimensional structure, which accounts for their individual functions. Beyond differential linkages (e.g. K48, K63, K11), LifeSensors has created subpanels of IQF diubiquitin substrates within each linkage.  Because each DUB is likely to recognize and cleave substrates with unique steric considerations, these subpanels vary in location of reporter fluorophore and quencher.  We recommend that each DUB be empirically evaluated against a panel of IQF DiUb substrates to select the the optimal fluorophore/quencher pairing.

IQF DiUb substrate panelsIn order to efficiently determine the optimal substrate for any DUB, or rapidly assess a panel of various DUBs, LifeSensors offers comprehensive panels for either K48 (DiUb48 panel cat no. DU0101) or K63 linked diubiquitins  (DiUb63 Panel DU0102). Each panel contain all six variants of the respective linkage along with a control enzyme (USP2core). In addition there is also a full (K48/K63) panel (DU0201) that contains all 12 variants.Need bulk quantities? We would be happy to work out a custom price that fits your budget. Please email info@lifesensors.com for more information.This product is covered by US patent # 8,518,660. By purchasing this product, the purchaser agrees to comply with the terms of our Limited Use Label License.

Info

Species	Human
Source	E. coli
Tag	None
Molecular Weight	18,478.3 +/- 18Da
Quantity	25 µL
Concentration	20 µM
Formulation	50 mM Sodium MES, pH 6.0
Storage	+4ºC

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。