Reliable ChIP primers for Sp1? - (Nov/17/2005 )
I try to find any primer sequence(s) that is reliable in ChIP. I did IP with Sp1 from Santa Cruz Biotech. However, I don't want to carry out all the way to the end to find out if the IP working or not (it's all cost effective and saving!). I want to check with Q-PCR if it's possible.
hi, I did chip for Sp1 with santa cruz antibody and used DHFR as a positive control... primers
As a positive control, a 68bp region of the DHFR (dihydrofolate reductase) promoter was amplified with the same reagents and the following primers: 5-TCGCCTGCACAAATAGGGAC-3 and 5-AGAACGCGCGGTCAAGTTT-3 using the following conditions for PCR: 2 min 94ºC, followed by 35 cycles of 94ºC for 30s, 57.4ºC for 30s, 72ºC for 30s, then one cycle of 72ºC for 10min.
Got the DHFR as positive from a paper on p53 where they used it for real time
Chromatin immunoprecipitation analysis fails to
support the latency model for regulation of
p53 DNA binding activity in vivo
M. D. Kaeser and R. D. Iggo*
Oncogene Group, Swiss Institute for Experimental Cancer Research (ISREC), 1066 Epalinges, Switzerland
Thanks very much, beccaf22. I will try out these primers on my samples (HL-60 cells).