Dendritic Cells and Flow Cytometry - (Apr/07/2009 )
I use 24 well plates. I have tried different cell concentrations like 2.5x10^5 to 1x10^6.
I used 5microgram/ml LPS based on some papers that I came across. I even emailed these people regarding the same and
they did not give me any constructive suggestions.
I will def try using low LPS concn and see how my cells behave.Will let u know.
Thanks for the suggestions.
Will bug you when I have more questions. 
-immunologist-
immunologist on Apr 29 2009, 01:14 PM said:
I use 24 well plates. I have tried different cell concentrations like 2.5x10^5 to 1x10^6.
I used 5microgram/ml LPS based on some papers that I came across. I even emailed these people regarding the same and
they did not give me any constructive suggestions.
I will def try using low LPS concn and see how my cells behave.Will let u know.
Thanks for the suggestions.
Will bug you when I have more questions.
I used 5microgram/ml LPS based on some papers that I came across. I even emailed these people regarding the same and
they did not give me any constructive suggestions.
I will def try using low LPS concn and see how my cells behave.Will let u know.
Thanks for the suggestions.
Will bug you when I have more questions.
Your procedure seems to be fine, you are using too much LPS - 10-20 ng/ml is sufficient. But I guess you are lacking the 2nd signal. Either TNF or IFN-gamma would provide the 2nd signal. I hope this helps. The Dc field is real convoluted...
-Spaced_out-