Editing Protein Production Enhancer for 293 Cells

HyClone SFM4HEK293 can substitute for the Invitrogen reagents with minimal loss of expression levels(10–20%).

[https://www.sciencedirect.com/science/article/pii/S1046202313002673 https://www.sciencedirect.com/science/article/pii/S1046202313002673]

<br/>cell cycle inhibitors, cyclin-dependent&nbsp;kinase inhibitor 1 (p21) and cyclin-dependent&nbsp;kinase inhibitor 1B (p27)

For enhanced expression, the antibody expression plasmid(s)<br/>were cotransfected with the p33SVLT plasmid, p21 and p27 at a<br/>weight ratio of 0.69/0.01/0.05/0.25 (antibody plasmid(s)/<br/>p33SVLT/pORF-hp21/ pORF-hp27).



A chemical inhibitor of caspases (caspase-3), z-VAD-fmk (Enzyme Systems Products),&nbsp; &nbsp;dissolve in DMSO to make a 100 mM (1000×) stock, add 4 ul to 60 mm dish containing 4 ml of culture 48 hours after transfection, and then add 4 ul every 24 hour. z-VAD-fmk has the effect of prolongation of cell viability.

https://resources.rndsystems.com/pdfs/datasheets/fmk004.pdf

Suramin protects against apoptosis during exponential growth, as indicated by the absence of DNA laddering and an increase in cell viability from roughly 70% to above 95%. Suramin also effectively dispersed cell aggregates so that single-cell suspension culture is possible. However, suramin does not protect against apoptosis during the death phase
[[Category:Antibodies]]