English
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.
Other Notes
MASMTGGQQMGRGHHHHHHGNLYFQGEVQVPEDPVVALVGTDATLCCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFAEGQDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYQGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSILRVVLGANGTYSCLVRNPVLQQDAHSSVTITPQRSPTGAVEVQVPEDPVVALVGTDATLRCSFSPEPGFSLAQLNLIWQLTDTKQLVHSFTEGRDQGSAYANRTALFPDLLAQGNASLRLQRVRVADEGSFTCFVSIRDFGSAAVSLQVAAPYSKPSMTLEPNKDLRPGDTVTITCSSYRGYPEAEVFWQDGQGVPLTGNVTTSQMANEQGLFDVHSVLRVVLGANGTYSCLVRNPVLQQDAHGSVTITGQPMTFPPEA
