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Material Data Sheet
His6-Rap80 UIM Domains
Price: $225.00
His6-Rap80 UIM Domains
Price:
$225
Quantity:
250 μg
Data Sheet:
Stock:
X mg/ml (X μM) 50 mM Hepes, 250 mM NaCl, 1 mM DTT pH 8.0
Purity:
>95% by SDS-PAGE Rap80 (Receptor Associated Protein 80) interacts with BRCA1, a ubiquitin E3-ligase which functions in conjugation with the BARD1 deubiquitinating enzyme. BRCA1 is recruited to DNA damage sites by poly-ubiquitin chains through Rap80 which contains 2 tandem ubiquitin-interacting motifs (UIMs). Rap80 constitutes a protein complex with ABRA1 which interacts with the BRCT domain of BRCA1. Upon DNA damage the Rap80-ABRA1 complex targets the BRCA1-BARD1 complex to K6- and K63-linked poly-Ub chains at these foci. The UIM domains of Rap80 have been shown to have preferential binding to K-6 and K63-linked Ub chains and binds to K48-chain with a much lower efficiency. It is not known if Rap80 UIMs interact with Ub chains linked via K11, K27 or K33. Rap80 has a low affinity for mono-, di- and tri-Ub but binds efficiently to tetra (or greater) ubiquitin chains. This protein can be used for the isolation and identification of ubiquitinated proteins that are non-K48-linked. This protein is His-tagged which allows for metal chelate affinity purification and also allows for convenient immuno-detection of conjugates using His6-specific antibodies.
Use & Storage
Use:
Use 50-100 μg of protein to pull down 10-20 μg of purified K63-linked ubiquitin chains. The amount necessary for use in crude lysates needs to be determined empirically.
Storage:
Store at -80°C. Avoid multiple freeze/thaw cycles. Literature
References:
Buchberger A. (2002) Tren. Cell. Biol. 12 :216-221
Hurley J.H., et al. (2006) Biochem. J. 399 :361-372
Kim H., et al. (2007) Science. 316:1202-1205
Kim H., et al. (2008) Mol. Cells. 25:457-61
Sobhian B., et al. (2007) Science. 316:1198-1202
Wang B., (2007) Science. 316:1194-1198
Wu W., et al. (2008) Cell. Div. 31-10
Yan J., et al. (2007) Canc. Res. 15:6647-6656






