Application

E-Cadherin has been used as coating matrix protein for 1) maintaining long-term ES or iPS cell culture then combine with proper ES cell culture media, 2) as a coating matrix material for 11R tag recombinant TF intracellular delivery for protein derived iPS protocol with extremely low-level non-specific interaction and 3) as a native antigen for antibody production.Use these recommendations as guidelines to determine the optimal coating conditions for your culture system. 1. Thaw E-Cadherin and dilute to desired concentration using serum-free medium or PBS. The final solution should be sufficiently dilute so that the volume added covers the surface evenly. Note: Use 1 ml PBS per well in a 6-well plate.2. Add 5 - 10 µg protein to each well and incubate at 2 to 10 °C overnight.3. After incubation, aspirate remaining material. 4. Plates are ready for use. They may also be stored at 2-8 °C damp or air dried if sterility is maintained.

Preparation Note

Recombinant human E-Cadherin gene (155-710 aa Fragment) was constructed with codon optimization and expressed in non-fusion protein form in E. coli as inclusion bodies. The final product was refolded using a unique "temperature shift inclusion body refolding" technology and chromatographically purified.

Sequence

MDWVIPPISCPENEKGPFPKNLVQIKSNKDKEGKVFYSITGQGADTPPVGVFIIERETGWLKVTEPLDRERIATYTLFSHAVSSNGNAVEDPMEILITVTDQNDNKPEFTQEVFKGSVMEGALPGTSVMEVTATDADDDVNTYNAAIAYTILSQDPELPDKNMFTINRNTGVISVVTTGLDRESFPTYTLVVQAADLQGEGLSTTATAVITVTDTNDNPPIFNPTTYKGQVPENEANVVITTLKVTDADAPNTPAWEAVYTILNDDGGQFVVTTNPVNNDGILKTAKGLDFEAKQQYILHVAVTNVVPFEVSLTTSTATVTVDVLDVNEAPIFVPPEKRVEVSEDFGVGQEITSYTAQEPDTFMEQKITYRIWRDTANWLEINPDTGAISTRAELDREDFEHVKNSTYTALIIATDNGSPVATGTGTLLLILSDVNDNAPIPEPRTIFFCERNPKPQVINIIDADLPPNTSPFTAELTHGASANWTIQYNDPTQESIILKPKMALEVGDYKINLKLMDNQNKDQVTTLEVSVCDCEGAAGVCRKAQPVEAGLQIP