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Thanks for your email and infor regarding freezing antibodies.
Here are basic infor about our ChIP procedures:
1. what dilution or concentration you're using for each antibody in ChIP?
We used 2 ug and 4 ug for 5 million cells in 1ml of ChIP binding buffer.
2. What kind of cells are you studying, and is the chromatin fresh?
U2OS cells, freshly corsslinked, used H3K36me3 as a positive control.
3. What genes are used as the positive controls?
We tested both total Pol II, S5P and S2P. We used Myc gene promoter 100bp, gene body 3K and 6K downstream of TSS as positive controls for them. At all the regions we only observed IgG levels of signals (0.0002-0.001% of Input). I was expecting 0.2-1% of input signals, based on our experience with the same Abs of previous lots.
Asked on Apr 25 2012
Thank you for sending the additional information, and I apologize for the extended delay getting back to you. Our ChIPscientist has been out of the office, but I have received further information from him regarding these antibodies.
I have found lots for each of these antibodies that will be suitable for your studies.
Please let me know if you approve of these, and I will send them out promptly. The vials of ab5131 and ab5408 will be sent overnight and delivered tomorrow, but the vial of ab5095 will take a few extra days to transfer from our UK office.
I apologize again for the delay, but I look forward to hearing from you. Please let me know if you have any further questions or if there is anything else that we can do for you.
Answered on Apr 25 2012