Retic Lysate IVT™ Kit, 60 Reactions - FAQs

When should Rabbit Reticulocyte Lysate or Wheat Germ Extract be chosen for in vitro translation reactions?

Rabbit Reticulocyte Lysate (RRL) and Wheat Germ Lysate (WGE) can often be used interchangeably for in vitro translation. In general, RRL efficiently translates proteins greater than 30 kDa while WGE works well for proteins of all sizes (including those in the range of 15-30 kDa), although larger proteins may be expressed more efficiently in RRL. This appears to be partially due to the relative instability of small proteins in RRL. However, certain templates are translated more efficiently in RRL, whereas others produce more protein in WGE; the difference in yield can be quite dramatic and must be determined empirically.

Another consideration is whether the protein of interest is present endogenously in either RRL or WGE, or whether factors that might interfere with downstream applications may be present in RRL or WGE. This can be determined by performing a control in vitro translation reaction with no template added. The RRL is recommended if Canine Pancreatic Microsomal Membranes are used for cotranslational processing (core glycosylation or signal peptide cleavage) or for production of proteins that require membrane insertion for in vitro translation.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.