How do you transfect Jurkat cells?
How do you transfect Jurkat cells?
Transfection of Jurkat Cells Plate 1 x 105 cells per well in 0.5 ml of complete growth medium. Cell density should be ~80% confluent on the day of transfection. For each well of cells to be transfected, dilute 0.5 μg of DNA into 100 μl of Opti-MEM® I Reduced Serum Medium without serum.
Are Jurkat cells hard to transfect?
However, presumably due to its T-cell origin, Jurkat cells are very difficult to transfect. Despite many previous efforts, non-viral vectors have not yet overcome the hurdles of low transfection efficiency and/or high toxicity in transfection of Jurkat cells.
How do you transfect HEK cells?
The protocol for a 24-well transfection reaction with HEK293 cells is here:
- Plate 10,000-15,000 HEK293 cells per well in 0.5 ml of complete growth medium 12-24 hours prior to transfection.
- Wash with 1xPBS and add 0.5 ml of fresh growth medium.
What are transient cells?
Transiently transfected cells express the foreign gene but do not integrate it into their genome. Thus the new gene will not be replicated. These cells express the transiently transfected gene for a finite period of time, usually several days, after which the foreign gene is lost through cell division or other factors.
Why are HEK cells easily transfect?
However, as a general rule the cell lines, like the HEK293, are easier to transfect than the primary cells. Most importantly is the cancerous nature of the cell lines make which makes them easier to accept foreign macro-molecules (DNA).
What is transient production?
Transient expression implies the temporary and small-scale production of antibodies in a mammalian host; while stable expression implies the stable, virtually everlasting, and large-scale production of recombinant proteins.
What is a transient cell line?
What is the difference between transient and stable transfection?
In stable transfection, the plasmid DNA successfully integrates into the cellular genome and will be passed on to future generations of the cell. However, in transient transfection, the transfected material enters the cell but does not get integrated into the cellular genome.