AB139472

E2-Ubiquitin Conjugation Kit

Name: E2-Ubiquitin Conjugation Kit (ab139472) | Abcam
Brand: Abcam Limited
SKU: AB139472
Price: 855 USD
Availability: InStock

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(6 Publications)

E2-Ubiquitin Conjugation Kit (ab139472) provides the tools necessary for generating a range of thioester-linked ubiquitin-conjugated E2 enzymes for use in ubiquitinylation experiments.

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Functional Studies - E2-Ubiquitin Conjugation Kit (AB139472)

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Supplier Data

Functional Studies - E2-Ubiquitin Conjugation Kit (AB139472)

Western Blot of Thioester Assays (TE +ve/-ve controls) for all E2 conjugating enzymes provided.

Biotinylated-ubiquitin-enzyme conjugates were detected by Western Blotting on thioester assays containing UbcH1 (A), UbcH2 (B), UbcH3 (C), UbcH5a (D), UbcH5b (E), UbcH5c (F), UbcH6 (G), UbcH7 (H), UbcH8 (I), UbcH10 (J), Ubc13/MMS2 (K) respectively, using Streptavidin-HRP detection system. M : Biotinylated SDS molecular weight marker, from bottom : 20.1, 29.0, 39.8, 58.1kDa.

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Western blot - E2-Ubiquitin Conjugation Kit (AB139472)

Western blot analysis of Thioester assays (TE (+)ve/(-)ve control) for E2 conjugating enzyme UbcH2. Procedure as described in the assay set up section (without DTT), and ubiquitin conjugate detected by streptavidin AP. The absence of conjugate in TE (-)ve control control demonstrate that formation of TE is ATP dependent (required for E1 activation).

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Key facts

Sample types

Purified protein

Assay type

Enzyme activity

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Product details

E2-Ubiquitin Conjugation Kit (ab139472) provides the tools necessary for generating a range of thioester-linked ubiquitin-conjugated E2 enzymes for use in ubiquitinylation experiments. Biotinylated ubiquitin (provided in the kit) can be detected with streptavidin-linked enzymes via SDS PAGE and western blotting.

This product provides sufficient material for a total of 50 reactions: 4 reactions with each included E2 enzyme.

Uses for this kit include:

1. Ubiquitinylation of target proteins in presence of dedicated E3 ligase. Panel of E2s provided for generation of E2-Ub thioester conjugates for testing vs. specific E3/target combinations. For example: ubiquitinylation of p53 in the presence of mdm2 (E3) and UbcH5b (E2).

2. Activation of Ub for thioester conjugation to novel E2 enzymes (substituted like for like with kit E2s, under directly comparable conditions).

Ubiquitinylation, the covalent attachment of ubiquitin to proteins, is achieved through three enzymatic steps. In an ATP-dependent process, the ubiquitin activating enzyme (E1) catalyzes the formation of a reactive thioester bond with ubiquitin, in the presence of a Mg2+ cofactor, followed by its subsequent transfer to the active site cysteine of a ubiquitin carrier protein (E2). The specificity of ubiquitin ligation arises from the subsequent association of the E2-ubiquitin thioester with a substrate specific ubiquitin-protein isopeptide ligase (E3), which facilitates the formation of the isopeptide linkage between ubiquitin and its target protein.

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What's included?

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Properties and storage information

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-80°C

Appropriate long-term storage conditions

-80°C

Storage information

-80°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Ubiquitin E2 also known as E2 ubiquitin-conjugating enzyme is essential in the ubiquitin-proteasome system mediating protein ubiquitination. It acts as a carrier transferring ubiquitin from E1 the ubiquitin-activating enzyme to E3 ligase the ubiquitin ligase facilitating the ubiquitin-conjugation process. Ubiquitin E2 proteins typically have molecular masses ranging from 14 kDa to 20 kDa. They distribute widely in eukaryotic cells playing roles across various tissues.

Biological function summary

Ubiquitin E2 enzymes contribute to protein degradation by tagging proteins for degradation by the 26S proteasome. They form a complex with E1 and E3 to ensure specific proteins are accurately marked for turnover. This precise marking regulates many cellular processes such as cell cycle progression DNA repair and response to oxidative stress.

Pathways

Ubiquitin E2 enzymes integrate into significant cellular pathways like the cell cycle and DNA damage response. They collaborate with other proteins such as cyclins and ubiquitin E3 ligases to regulate cell division. During DNA repair ubiquitin E2 works with RAD proteins to identify and process damaged DNA maintaining genomic stability.

Ubiquitin E2 enzymes associate with cancer and neurodegenerative diseases such as Parkinson's disease. Their dysregulation leads to abnormal protein accumulation or degradation contributing to disease pathogenesis. They interact with proteins like Parkin an E3 ligase which is significant in Parkinson's disease and p53 a tumor suppressor protein highlighting their importance in these disorders.

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Product protocols

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Target data

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Publications (6)

Recent publications for all applications. Explore the full list and refine your search

Nature cell biology 24:723-736 PubMed35469017

2022

CDK1-cyclin-B1-induced kindlin degradation drives focal adhesion disassembly at mitotic entry.

Applications

Unspecified application

Species

Unspecified reactive species

Nan-Peng Chen,Jonas Aretz,Reinhard Fässler

Open biology 11:210221 PubMed34610268

2021

BRCA1-BARD1 regulates transcription through modulating topoisomerase IIβ.

Applications

Unspecified application

Species

Unspecified reactive species

Heeyoun Bunch,Jaehyeon Jeong,Keunsoo Kang,Doo Sin Jo,Anh T Q Cong,Deukyeong Kim,Donguk Kim,Dong-Hyung Cho,You Mie Lee,Benjamin P C Chen,Matthew J Schellenberg,Stuart K Calderwood

Autophagy 17:3030-3047 PubMed33280498

2020

RNF186 regulates EFNB1 (ephrin B1)-EPHB2-induced autophagy in the colonic epithelial cells for the maintenance of intestinal homeostasis.

Applications

Unspecified application

Species

Unspecified reactive species

Huazhi Zhang,Zhihui Cui,Du Cheng,Yanyun Du,Xiaoli Guo,Ru Gao,Jianwen Chen,Wanwei Sun,Ruirui He,Xiaojian Ma,Qianwen Peng,Bradley N Martin,Wei Yan,Yueguang Rong,Chenhui Wang

Scientific reports 8:3471 PubMed29472632

2018

African swine fever virus encodes for an E2-ubiquitin conjugating enzyme that is mono- and di-ubiquitinated and required for viral replication cycle.

Applications

Unspecified application

Species

Unspecified reactive species

Ferdinando B Freitas,Gonçalo Frouco,Carlos Martins,Fernando Ferreira

Science signaling 9:ra78 PubMed27485016

2016

Small heterodimer partner SHP mediates liver X receptor (LXR)-dependent suppression of inflammatory signaling by promoting LXR SUMOylation specifically in astrocytes.

Applications

Unspecified application

Species

Unspecified reactive species

Jee Hoon Lee,Hyunmi Kim,Soo Jung Park,Joo Hong Woo,Eun-Hye Joe,Ilo Jou

Scientific reports 5:14715 PubMed26423026

2015

Natural small molecule FMHM inhibits lipopolysaccharide-induced inflammatory response by promoting TRAF6 degradation via K48-linked polyubiquitination.

Applications

Unspecified application

Species

Unspecified reactive species

Ke-Wu Zeng,Li-Xi Liao,Hai-Ning Lv,Fang-Jiao Song,Qian Yu,Xin Dong,Jun Li,Yong Jiang,Peng-Fei Tu

View all publications

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Product promise

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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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E2-Ubiquitin Conjugation Kit

Functional Studies - E2-Ubiquitin Conjugation Kit (AB139472)

Western blot - E2-Ubiquitin Conjugation Kit (AB139472)

Key facts

Sample types

Assay type

Product details

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (6)

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