Cellular ELISA - (Apr/10/2007 )

Hi ,
I want to do cellular ELISA using dendritic cells.After coating the plates with specific cell concentration ,how can I determine how many cells have actually adhered to the plate?Do I just go ahead and use the plates assuming that all the cells have adhered or incorporate a control to determine how many cells have actually adhered to the plate?
Any help will be appreciated.

-Seema Haridas-

QUOTE (Seema Haridas @ Apr 11 2007, 05:04 AM)

Why is the specific cell concentration essential? What concentration are you working with? Can you collect the liquid from the binding step, and count the cells remaining? A haemocytometer will enable you to get a reasonably accurate count of what's left.

-swanny-

Yeah..that could be done.Did not think about it.
Thanks..

-Seema Haridas-

Hi, I have a related question:

how do I develope an ELISA protocol for measuring the concentration of a mAb for a cell surface marker (such as CD4 or CD8)?
with what do I do the coating on the plate?

Does anybody have a suggestion?

-taliasa-

QUOTE (taliasa @ May 16 2007, 10:41 PM)

Assuming you have purified the mAb, why not just do a protein concentration assay? You could use a known IgG, say, for t he standard curve.
If you have to do an ELISA, again assuming your mAb is pure, you could coat with soluble CD4 / CD8 / whatever.

-swanny-