KIT UBI trap-B for Western-blot

UBI traps are used to isolate and identify ubiquitylated proteins from cells, tissues or organs. Multiple ubiquitin binding domains (UBDs) contribute to enhance their affinity for polyubiquitin chains compared to single UBD. (1-3). Prototype UBI traps interfere with the action of de-ubiquitylating enzymes (DUBs) and the proteasome which contribute to increase the yield of ubiquitylated proteins purified and facilitate the detection of low abundance proteins. These unique properties are not observed with any other technologies that can be used in a large panel of applications (3).
UBI trap-B is based on UBDs with preference for K63 linked polyubiquitin chains but other chains can also be captured (1-3). Furthermore, differences can be observed depending on the relative chain abundance in distinct cell lines and response to various treatments. GST control is provided for a single control reaction. KIT UBI trap-B contains all reagents and solutions required for the capture of Ubiquitylated proteins from cell lines, tissues and organs (see below).

>30 mg/mL

RT but once reconstituted store at -80°C and avoid freeze/thaw cycles

Higher affinity for polyubiquitin chains than a single UBD
Avoid overexpression of tagged ubiquitin for pull downs
Protect poly-ubiquitin chains without inhibitors specific to the proteasome or DUBs activity
Replace ubiquitin specific antibodies for enrichment of poly-ubiquitylated proteins
KIT UBI trap-B has been developed to facilitate the use, long-term stability and transport of all components required to capture ubiquitylated proteins using agarose beads.

UBI trap-B is not crosslinked to agarose beads but preserved in a lyophilized state for long term storage. UBI trap-B is reconstituted with a specific buffer just before use and mixed with glutathione agarose beads provided in the kit. Washing and elution buffers are also included. Negative control GST is provided for a single control reaction. Captured proteins have to be boiled to be analyzed by Western blot. Since UBI traps are not crosslinked to agarose, these will be released during the boiling. Avoid using antibodies for Western blot analysis that have been generated with a GST fusion with the protein of interest, otherwise GST protein will be detected. If no alternative antibodies are available, we advise to use UBI traps tagged with biotin.