Application

Coating a plate well (6 well plate) with this recombinant CD276 protein in T cell specific medium at 1-10 µg/well allows for use 1) for human T cell / receptor interaction study in vitro or 2) as a highly purified recombinant antigen as cancer biomarker for diagnosis application development.Use this procedure as a guideline to determine optimal coating conditions for the culture system of choice.1. Thaw CD276 and dilute to desired concentration using serum-free medium or PBS. The final solution should be sufficiently dilute so the volume added covers the surface evenly (1-10 µg/well, 6 well plate).Note: Use 1 ml PBS per well in a 6-well plate. 2. Add 1 - 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

The full-length extracellular domain of the human CD276 gene (29 - 466 aa) was constructed with 29 N-terminal T7/HIS-tag and expressed in E. coli as inclusion bodies. The final product was refolded using a unique “temperature shift inclusion body refolding” technology and chromatographically purified.

Sequence

MASMTGGQQMGRGHHHHHHGNLYFQGEVQVPEDPVVALVGTDATLCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSILRVVLGANGTYSCLVRNPVLQQDAHSSVTITPQRSPTGAVEVQVPEDPVVALVGTDATLRCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFTEGRDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYRGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSVLRVVLGANGTYSCLVRNPVLQQDAHGSVTITGQPMTFPPEA