Pan-branch Ubiquitin TUBE-UBQLN1 (Magnetic Bead Conjugate) #43658
- IP
Product Information
Product Description
Product Usage Information
Add beads at a 1:20 dilution to cell lysates prepared in 1X Cell Lysis Buffer (10X) #9803. Mix overnight at 4°C. Wash five times in 1X Cell Lysis Buffer. For more details, refer to the Immunoprecipitation for Native Proteins protocol.
For bead washing and subsequent washing of affinity-purified complexes, the beads can be separated from solution using our 6-Tube Magnetic Separation Rack #7017. Place the tubes containing the beads in the Magnetic Separation Rack and wait 1 to 2 min for solution to clear before carefully removing the supernatant. Remove the tubes from the Magnetic Separation Rack, add new solution, and resuspend the beads by gently vortexing or rocking the tubes.
Storage
Protocol
Specificity / Sensitivity
Species Reactivity:
Source / Purification
Background
Substrate proteins are linked to ubiquitin using seven distinct ubiquitin lysine residues (Lys6, Lys11, Lys27, Lys29, Lys33, Lys48, and Lys63). Formation of a polyubiquitin chain occurs when a lysine residue of ubiquitin is linked to the carboxy-terminal glycine of another ubiquitin. Proteins polyubiquitinated at specific lysine residues display a tendency to be targeted for different processes; K48-linked polyubiquitin chains mainly target proteins for proteasomal degradation, while K63-linked polyubiquitin chains regulate protein function, subcellular localization, or protein-protein interactions (8). K63-linked polyubiquitin chains exert nonproteolytic functions in vivo, such as protein trafficking, kinase/phosphatase activation, and DNA damage control, all of which might be important in regulation of cancer survival and development (9,10).
Ubiquitin-associated (UBA) domains are protein regions that interact with ubiquitin. Tandem-repeated ubiquitin-binding entities (TUBEs) were designed by using four tandem UBA domains, based on the theory that tetraubiquitin chains are a minimum requirement for efficient proteasomal degradation (11). TUBEs designed with UBA domains from UBQLN1 and RAD23A bind to K48- and K63-linked tetraubiquitin chains and can be used to efficiently purify ubiquitylated proteins from cell extracts (12).
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- Jesenberger, V. and Jentsch, S. (2002) Nat Rev Mol Cell Biol 3, 112-21.
- Komander, D. (2009) Biochem Soc Trans 37, 937-53.
- Chen, Z.J. and Sun, L.J. (2009) Mol Cell 33, 275-86.
- Yang, W.L. et al. (2010) Oncogene 29, 4493-503.
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- Hjerpe, R. et al. (2009) EMBO Rep 10, 1250-8.
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