Jun 9, 2008 · I am working with HEK 293T cells, and I am having some serious problems getting them to attach to the culture flasks and dishes i use. There seems to be no pattern in the way they behave, sometimes they attach and grow and sometimes they don't. It would seem they prefere NUNC products, but not always.

Apr 8, 2010 · How many 293T or Hela cells should I seed for transfection in 10cm dishes (I want to harvest after 48h)? And something else just came to my mind: I know that if you have a selective medium with antibiotics (puromycin, g418 etc.), you shouldn't use the antibiotics in the medium when you are seeding cells for transfection.

Sep 14, 2005 · 293 and HEK293 cells are same 293T and HEK293T cells are same 293T cells are 293 cells transformed by expression of the large T antigen from SV40 virus that inactivates pRb. you can get info on large T here Fred

Many floating cells. This experiment has repeated with other 293T stock and showed that 2 million starting seed is not good for cell attachment. and I will have to wait more time for it to grow into confluency. But, I've been growing 293T for the past 6 months, and they always grow and attach well even I put 1 million starting cells during passage.

Sep 26, 2006 · I have been working with HEK293 cells for a while and recently have been trying to generate stable cells expressing my protein of interest, unfortunately without success (using G418 selection). I have obtained transient expression of my protein of interest and have also been able to generate cells that stably express a fluorescent protein alone.

I was wondering if anyone can tell me whether HEK293T cells are resistant to Geneticin (G418) in general. I know you can make them reisistant to Geneticin by transfection of a neomycin resistant plasmid but I want to know if they are already resistant before I start.

My 293T cells - actually a cell line which is known to be transfected very easily and efficiently (no matter by which method) - behave quite strange because control transfection with GFP e.g. look nice under the microscope + on Western Blots but my …

If the PBS wash is done for a few seconds, you will not lose cells. But if you to wash several times, use PBS with calcium and magnesium. It can help. and iif you still have a problem, coat the dishes or plates with poly lysine and then seed the cells. This can help cell attachment.

Dec 1, 2009 · I've been trying to transiently transfect U2OS cells to carry out co-IPs using lipofectin/lipofectamine reagents. I've worked with 293T cells and I managed to always get good levels of transfection, without having to worry too much about cell density or reagent:DNA ratio, but with the U2OS I can't seem to be able to establish a procedure that ...

I did an experiment with 293T cells, which I transiently transfected at 60-70% confluency. I did not do any cellcounts, but now i need to know (by estimation) how many cells I had at the time of the transfection and how many cells I had 24h posttransfection. I used 24 well plates but if anybody knows this for 10 cm dishes I can calculate from ...