how many antibody eveyone use for CHIP assay - (May/15/2006 )

I have done several times CHIP asaay but i cann't get enough DNA for PCR. everytime i used one flask of 150cm of cells but i can just got 1-2 ng DNA finally for PCR. the antibody i used for each assay is around 50ul, and because the antibody of upstate didn't show concentration one the data sheet,i can not know if i used too much or too little make make me cann't get what i want.
Would the anyone help me handle this poblem? Thanks a lot

Hi,

I haven't had this problem so far. I always use 10cm dishes and 3 micrograms of antibody and in the last step I use the DNeasy Tissue Kit from Qiagen to isolate my DNA. I get my DNA in a volume of 200 microliters and I use 2,5 for Real-time.

Cheers

I harvest MEFs cells from 8 dishes, about 2.4x107 cells. I use upstate antibody 10 ul for each sample.