This kit allows you to detect whether your target protein is ubiquitylated and determine the linkage type of the ubiquitin chain.
Determining the linkage of polyubiquitin on target proteins is challenging. The traditional methods are either through Mass Spectrometry or immunoblot with
linkage specific antibodies, which are cumbersome. LifeSensors has developed a more definitive method for demonstrating the ubiquitylation linkage of a protein, which is to couple immunoprecipitation of polyubiquitylated protein with digestion by a linkage specific deubiquitylase prior to immunoblot analysis. An increased signal for the unmodified substrate or a decreased signal of polyubiquitylated substrate at high molecular weight after K48/K63 specific DUB treatment is a clear indication that the protein was K48/K63 ubiquitylated. We have built this kit around the use of Tandem Ubiquitin Binding Entities (TUBEs) which bind to all ubiquitin chain linkages.
This kit includes the following DUBs:
- A Lys48 linkage specific DUB - 25 µg, 10 µM
- A Lys63 linkage specific DUB - 35 µg, 10 µM
This product is protected by one or more US or Foreign
- A broad spectrum DUB - 25 µg, 10 µM
patents. Please read the Limited Use Label License to learn more. By purchasing this product, the purchaser agrees to comply with these terms.
- Convenient one-step pull-down of polyubiquitylated proteins from cell and tissue extracts
- Isolate ubiquitylated proteins for proteomic studies
- Confirmation of target protein ubiquitylation
- Real proof of ubiquitylation – no mass spec needed!
- Verification that the pattern observed is from ubiquitin, not other post-translational modifications
- Easy comparison of polyubiquitin levels between treated samples
- Easy screening for multiple POIs
- No substrate purification needed
- Control for antibody not recognizing ubiquitylated substrate
- Sufficient for a minimum of 10 pull-downs
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