How to treat cells with TNFa - (Nov/20/2012 )

Hi All ,

I need to know what is the procedure for each of the following:-
How to treat cells with TNFa 
How to treat cells with OPG (Osteoprotegerin)

I am using glutamix-DMEM medium 

Finally , I have 50micrograms/ml TNFa (aliquot)
How can I get 10ng/ml (aliquot)


Thank u all.

For the first two questions, I would suggest looking up papers where researchers have done this (preferably with a cell line similar to yours) as a starting point.

For the second part, have a go at figuring it out and post that up- then we can help you from there. It is an essential skill to be able to make solutions at concentrations you need, so best to learn it now rather than us just giving you the answer and you needing help again the next time you have to do it

As a starting point, I suggest converting the concentrations to the same units. It will make it easier.

(so 50ug/ml is actually 50 000 ng/ml)

Zerro on Wed Nov 21 03:23:07 2012 said:

Dear Zerro,

I have used TNF alpha many times and here are my tips:-

Reconstitute the lyophilised TNF alpha in complete media i.e. with foetal calf serum. Some poeple just use PBS with out any added protein and this is a big mistake.

Dilute the TNF alpha into "working aliquots" i.e. a working concentration that is relevant to your experiments. The stock or working solution should be easily diluted.

The aliquots should be stored at -80 oC in my experience if you are to keep them for longer than a couple of months.


Lastly, I completely agree with leelee (as I always do), have a go at the dilution yourself. I will however give you a bigger clue:-

As leelee states: "so 50ug/ml is actually 50,000ng/ml"

I will go a little further: so 50,000ng/1000ul = 50ug/ml and a 1:10 Dilution is 1ml to 9ml of diluent

Hope this is useful, have fun trying

Kindest regards

Uncle Rhombus