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Home > Protein Expression > Mammalian expression

Mammalian expression

HongTide can provide you mammalian protein expression service with the produciton ability from milligram to grams. Density of cell culture can reach to 2-4×106 cells/ml. Our mammalian protein expression service includes transient transfection and screening of stable cell lines.
Service Procedures and Specification
1. Programme analysis
Timeline
Deliverables

  • Optimization of codon

  • Fusion of tag

  • Purification methods

  • Optional host cells

1-2 days

  • Programme analysis report

2. Expression system selection
Optional expression vector
pMZ-X3 related series carrier (secretory type, optional His-tag)
Other commercial carriers
Optional expression host
HEK293 series cells CHO series cells (adherent or suspension cells)
Other commercial strains
3. Expression vector construction
Timeline
Deliverables
A. Gene optimization and gene synthesis
B. Subcloning into eukaryotic expression vector
C. Endotoxin-free plasmid extraction
Gene synthesis: about 2 weeks
Subcloning: 1-2 weeks
1. Gene synthesis report
2. Sequencing verification report
3. E.coli DH10 containing expression plasmid
4. Expression plasmid
4. Transient transfection test
Timeline
Deliverables
D. Cell transient transfection
E. Express test
F. Expression analysis & identification(WB)
Cell transfection: 1 week
Expression identification: 1-2 weeks
Expression analysis report
5. Expression and purification
Timeline
Deliverables
G. The optimal condition is 200ml
H. Ni column affinity purification
I. Recombinant protein identification
Expression purification assay: 1-2 weeks
Protein sample (50-500ug protein available)
6. Amplification and protein purification
Timeline
Deliverables
J. Amplification(according to specific needs)
K. Ni column affinity purification
L. Purity identification (PAGE)
M. Protein dispensing
Amplification: 2-3 weeks
Expression purification: 1-2 weeks
1. Protein purity identification report
2. Finished protein
(The specific amount is based on the level of the small test expression)

Supplementary explanation

1. Expression levels of different proteins vary widely, we will deliver aimed recombinant proteins as many as possible during the pilot phase.
2. Usually we use Mouse-Anti-His antibody to detect recombinant protein. If you want to use antibody with no tag, please provide specific antibody for WB detection.
3. If you want your protein without any tags, we will use AKATA protein purification equipment to separate the protein, but the purification cost will increase.
4. We will provide a full set of experimental procedures and original photos even if we cannot obtain the target protein due to the low expression or no pression of protein.