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
| biological source | human |
| recombinant | expressed in E. coli |
| description | 0.1 mg recombinant human CD276 in 20 mM Tris-HCl buffer, containing NaCl, KCl, EDTA, L-arginine, DTT and glycerol. |
| sterility | Filtered sterilized solution |
| assay | ≥90% (SDS-PAGE) |
| form | liquid |
| packaging | pkg of 100 μg |
| concentration | 0.5 mg protein/mL |
| technique(s) | cell culture | mammalian: suitable |
| accession no. | NP_001019907 |
| shipped in | dry ice |
| storage temp. | −20°C |
| Gene Information | human ... CD276(80381) |
