PurMaFectin™ 293T System

Description

Background

PurMaFectin™ 293T System provides exceptional transient and stable transfection with over 95% efficacy.  Moreover, it offers an exceptional result for commercial cell lines including HEK 293, COS1, COS9 cells and adhesive cell lines. It completes the procedure in 35-40 minutes.

PurMaFectin™ 293T System Special Features

• It is easy to use.
• PurMaFectin™ 293T System does not cause cell toxicity.
• Provides consistent and reproducible transfection.
• More importantly, this transfection reagent suits both transient and stable transfection.
• PurMaFectin™ 293T protects DNA from lysosomal degradation and facilitates efficient plasmid delivery into the eukaryotic nucleus.
• Also, it provides effective, reproducible, and affordable benefits for scientific research.

Brief Guideline

Below, you can find some brief guidelines. For detailed protocol, click here:PurMaFectin293 User Manual

• For the highest efficacy and the lowest dead cells, transfect cells at around 70~80% confluency. Additionally, we strongly recommend starting the seeding by counting trypsinized cells so you maintain high consistency among your experiments.
• Different cell types react differently to the number of passages when they are used for transfection. To minimize the impact of passage on efficiency of transfection, either use the cells with the same passage number or use at least two different concentrations of transfection reagent as control in new transfect experiments before you optimize the condition.
• For HeLa cells, high-level transfection requires the presence of serum in the medium during transfection. The amount of required serum depends on sequence and the length of plasmid. The exact level of serum should be determined empirically.
• Furthermore, we deplete endotoxin from our transfection reagent. The endotoxin-contaminated DNA results in inefficient transfection and can cause high cellular toxicity. To deplete the endotoxin, we use PurMa^TM Endotoxin Elimination Column (Cat# P6H1216251). In this proprietary kit, Sepharose 6B covalently conjugates to an endotoxin-specific absorbent. The mentioned procedure will end with passing the reagent through 0.1µm filters 3X (included in the kit). The mentioned procedure eliminates the residual endotoxin.

References

1. A Guide to Transient Expression of Membrane Proteins in HEK-293 Cells for Functional Characterization. Ooi et al. Front Physiol. 2016 Jul 19; 7:300. doi: 10.3389/fphys .2016 .00300. eCollection 2016. PMID: 27486406.
2. Gene delivery in adherent and suspension cells using the combined physical methods. Kardani et al. Cytotechnology. 2022 Apr;74(2):245-257. doi: 10.1007/s10616-022-00524-4. Epub 2022 Feb 3. PMID: 35464169.