...optimized is the tobacco etch virus (TEV) protease. A TEV protease cleavage site is frequently placed between... 2: Protease recognition sites commonly used with tags Protease Recognition site Notes TEV ENLYFQS...reaction taking place between column runs. The TEV protease itself, with various mutations used to increase...purification, detection, solubilization, localization, or protease protection. Thus far Plasmids 101 has covered ...comprised of a calmodulin binding peptide (CBP), a TEV cleavage site (more on that in a moment), and 2 ProtA... that purpose has been served. At this point, proteases can be your friend rather your enemy. Two common... tags (SUMO and FLAG) are cleaved by specific proteases without requiring the addition of an independent...

... connected to a TEV protease cleavage site. TEV protease is a highly specific protease that is commonly... These signals activate a cascade of caspases, protease enzymes that cleave proteins. Executioner caspases...effectively study apoptosis in vivo scientists need a protease reporter that has a large dynamic range and high... GFP11) to increase fluorescence 10-fold after protease activity (To et al., 2016). However, the lab wanted...with a system where fluorescence was dependent on protease activation. Normally, β10-11 are antiparallel....position and fluoresce. FlipCherry: Red fluorescence protease reporter To add to the fluorogenic reporter toolkit...toolkit, the Shu Lab also designed a red fluorogenic protease reporter based on superfolder Cherry (sfCherry...

...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor...efficiency of tobacco etch virus protease (TEV), a highly sequence-specific protease used to cleave affinity tags...of the TEV cleavage and improve utility of the system for use with low affinity variants of TEV. The new...extracellular surface) which enables tracking of protease activity with a fluorescent protein--no affinity...

...modules with each possible combination of protease site (TEV or PreScission), epitope tag (12xHis, 2xStrep...plasmids, combining fluorescent tags, epitope tags, protease sites, and selection markers. These are especially...collection will yield an insert with the format (protease site)-6xGly linker-(epitope tag)-ADH1 terminator...

...Addgene ID Plasmid Gene/insert 165509 pOpen-TEV protease TEV protease 165565 pOpen-RNAse A RNAse A 165575 pOpen-RNAse...

...beta-arrestin-TEV protease fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription...

...disulfide bonds fold properly. If a TEV protease site is present, TEV can remove these tags from the purified...

...few of our proteins that lacked TEV. Although we were unable to cleave TEV within the protein we tested,...bacterial extract. The MBP tag can be removed using Xa protease. The carboxyl-terminus of the fusion protein includes...the AviTag. As a side note, we found that adding a TEV linker dramatically increased accessibility of the...

...Gitler 29629 GST-TEV-FUS FUS TEV, GST T7 ALS Aaron Gitler 29630 GST-TEV-FUS-267-526aa FUS TEV, GST T7 ALS ...29631 GST-TEV-FUS-1-373aa FUS TEV, GST T7 ALS Aaron Gitler 29632 GST-TEV-FUS-1-413aa FUS TEV, GST T7 ALS...pGST2-GST-TEV-OPTN S177D S473D OPTN GST-TEV ALS Sascha Martens 187830 pFastBac_Dual_GST-TEV-EGFP-TBK1 ...pET-Duet1-His-TEV-mCherry-OPTN WT OPTN His, TEV, mCherry ALS Sascha Martens 190192 pET-Duet1-6xHis-TEV-OPTN OPTN...-GST-TEV-mCherry-TBK1 TBK1 mCherry, GST, Tev P10 ALS Sascha Martens 198037 pFastBac_Dual_GST-TEV-EGFP-...666) TBK1 GFP, GST, Tev P10 ALS Sascha Martens 198428 pGST2-GST-TEV-OPTN OPTN TEV, GST tac ALS Sascha ...PRKCG PRKCG His, TEV T7 Spinocerebellar ataxia 18 Nicola Burgess-Brown 39042 GAK GAK TEV, His T7 Parkinson's...

... low protein yield, can be reduced by adding a protease inhibitor cocktail and phosphatase inhibitors ...for a longer time Protein degradation Add protease inhibitors to lysate Insufficient denaturing...or place a double ladder on the lefthand side. Whatever you do, be consistent and document it.   Did it...