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5126 Sigma-Aldrich

CD274 human

recombinant, expressed in E. coli, 0.5 mg protein/mL

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Properties

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biological source   human
recombinant   expressed in E. coli
description   0.1 mg recombinant human CD274 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
suitability   suitable for cell culture
accession no.   NP_054862.1
storage temp.   −20°C

Description

Preparation Note

The extracellular domain of recombinant human CD274 (19-238 aa, derived from BC074984) was constructed with codon optimization and expressed with a small T7-His-TEV cleavage site Tag (29aa) fusion at its N-terminal and expressed in E. coli as inclusion bodies. The final product was refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified.

Application

Coating a plate well (6 well plate) with this recombinant CD274 protein in a specific culture medium at 5-10 μg/well allows for use 1) as human T and B cells activation/differentiation studies or 2) as a potential biomarker protein for infectious diseases in vitro or 3) for auto-immuno disease diagnostic development.

Use this procedure as a guideline to determine optimal coating conditions for the culture system of choice.
1. Thaw CD274 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 (5-10 μg/well, 6 well plate).
2. Add appropriate amount of diluted material to culture surface.
3. Incubate at room temperature for approximately 1.5 hours.
4. Aspirate remaining material.
5. Rinse plates carefully with water and avoid scratching bottom surface of plates.
6. Plates are ready for use. They may also be stored at 2-8 °C damp or air dried if sterility is maintained.

Sequence

MASMTGGQQMGRGHHHHHHGNLYFQG^GEFFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNER

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
2

Documents

Certificate of Analysis


Cell Culture Guide

Milli-Q® Water Purification Solutions
Protocols & Articles

Articles

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Peer-Reviewed Papers
15

References

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