|Product Name:||TVMV C4 Protease|
|Manufacture:||LD Biopharma, Inc.|
|Intruduction:||TVMV protease derived from Tobacco Vein Mottling Virus, recognizes a linear epitope of the general form T_V_R_F_Q / S with cleavage occurring between Q and S. The protease is used to cleave affinity tag from fusion protein. The optimal temperature for cleavage is 30 °C. TVMV protease could be easily removed from the cleavage reaction by affinity chromatography using the polyhistidine tag at N-terminal of the TVMV C4 enzyme.|
|Gene Symbol:||TVMV C4 Protease|
|Species:||Tobacco Vein Mottling Virus (TVMV)|
|Package Size:||50 µg / Vial|
|Composition:||1.0 mg/ml in PBS with 20% Glycerol.|
|Storage:||Keep at -80°C for long term storage. Product is stable at 4 °C for 2-3 weeks.|
|Key Reference:||Lutz, R. and Bujard, H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucl. Acids Res. 25: 1203-10 (1997)
Yoon, H. Y., Hwang, D. C., Choi, K. Y., and Song, B. D. Proteolytic processing of oligopeptides containing the target sequences by the recombinant tobacco vein mottling virus NIa protease. Mol. Cells 10: 213-219 (2000)
Typical Reaction Condition
The "standard" reaction buffer for TVMV C4 protease is 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA and 1mM DTT. The duration of the cleavage reaction is typically overnight, although lots of cleavage will happen in the first few hours and prolonged incubation times may not lead to proportional increases in cleavage. TVMV C4 protease is maximally active at 34 °C, but we recommend performing the digest at room temperature (20 °C) or 4 °C overnight.
Typically, a good rule of thumb for initial test of digestion is 1 ug C4 enzyme per 50 – 100 ug target protein ratio. Perform a small-scale reaction first, if possible, to gauge the efficiency of processing.
1. Purity: > 95% by SDS-PAGE.
2. Activity: Completely digestion 50ug of MBP-IL6 fusion protein using 1 ug C4 enzyme (in 1 : 50 ratio) for 12 hours at 4C.
|Recombinant Protein Sequence:||MASMTGGQQMGRGHHHHHHGNLYFQGGEFSKALLKGVRDFNPISACVWLLENSSDGHSERLFGIGFGPYIIANQHLFRRNNGELTIKTMHGEFKVKNSTQLQMKPVEGRDIIVIKMAKDFPPFPQKLKFRQPTIKDRVCMVSTNFQQKSVSSLVSESSHIVHKEDTSFWQHWITTKDGQCGSPLVSIIDGNILGIHSLTHTTNGSNYFVEFPEKFVATYLDAADGWCKNWKFNADKISWGSFTLVEDAPEDDFMAKKTVAAIMD