PheBo Fusion Protein Cloning
A complete system for fusion cloning, cleavage, and purification



APPLICATIONS

In the PheBo-System for fusion protein cloning and purification, the protein of interest is cloned next to the leader protein ß-lactamase. The expressed fusion protein remains in the cytoplasm (vectors pHKcyto1 and pHKcyto2) or is released into the periplasm with the help of a signal sequence (vectors pHKperi1 and pHKperi2). ß-lactamase allows efficient protein purification under mild conditions on pre-packed re-usable Phenylboronate (PheBo-) affinity columns. The leader protein is removed by the site-specific Endoproteinase Pro-Pro-Y-Pro (Igase), which also cleaves inclusion bodies. After a second passage through the same affinity column, the protein of interest can be obtained in pure form from the column flow-through, whereas ß-lactamase and the endoproteinase remain on the column.


References:
  1. Kolmar, K. et al., J.Mol.Biol. 228 (1992) 359-365.
  2. Frisch. C, et al., Biol.Chem.Hoppe-Seyler 375 (1994) 353-356.


ORDER INFORMATION
Cat. No.DescriptionSize/Vol.
Kits with 1 vector (5 µg), Endoproteinase Pro-Pro-Y-Pro (50 µg), 2 ml PheBo Column:
5300-500pHKperi1
5300-510pHKperi2
5300-511pHKcyto1
5300-512pHKcyto2
Kits with 2 vectors (5 µg each), Endoproteinase Pro-Pro-Y-Pro (50 µg), 2 ml PheBo Column:
5300-513pHKperi1,pHKperi2
5300-514pHKcyto1,pHKcyto2
Kit with all 4 vectors (5 µg each), Endoproteinase Pro-Pro-Y-Pro (50 µg), 2 ml PheBo Column:
5300-515KIT
5300-516Endoproteinase Pro-PRo-Y-Pro (Igase)50 µg
5300-517Endoproteinase Pro-PRo-Y-Pro (Igase)250 µg
5300-5182 ml Phenylboronate Column
5300-5195 ml Phenylboronate Column
Vectors can be purchased separately. Vectors and columns shipped at RT; store at 4°C.
Proteases are shipped at 4°C; store at -20°C.
A detailed handbook is provided.





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