Hi Verma to do ADCC you have to have a either macrophages, neutrophils, NK cells as cytotoxic cells and a target that is recognized by the antibody so if you have a general IgG it would not work, there is no real target. Besides the binging of TIRC could hamper the binding to the Fc receptor. Thus, start form finding an antobody that recognizes a target in the cell mebrane of the target cell, if possible from different species, and then use PBMC at different ratios 1 target:5 effector, 1:10, 1:20 and 1:40

good luck 

Hi Juan,

I have read that for antibody binding to PBMC, target cell is not required. It is required only for the release of lytic components from the effector cells.

So, my IgG should bind to the PBMC (this I confirmed by visualizing my IgG bound PBMC under fluorescence microscope).

Also, I have an antigen against my IgG to which I m trying to bind it. 

Then Verma use the antigen to a solid phase, bing the antidody to the antigen and see if the cells bind to it before doing the ADCC, just a proposition

regards,

Thanks Juan,

I have tried that as well. Didn't work!

Another option is to use sheep erythrocytes to see binding of PBMC, T and NK cells mainly. The rosette should be your control of cell binding which I think is your main problem

regards

Will try that.

Thanks.