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Old 01-27-2012, 08:50 PM   #1
osl
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Default Good anti-GFP/YFP for ChIP-Seq?

Hi all,
Just wondering if anyone could recommend a good anti-GFP/YFP for ChIP-Seq.
I've tried quite a few (including ab290, JL-8) for ChIP-qPCR. I just found the polyclonal gives rather high background, and the monoclonal seems to have low efficiency (I'm working on Arabidopsis).
Also, for ChIP-Seq, is monoclonal Ab a better choice over polyclonal?
Thanks ahead.
os
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Old 01-27-2012, 10:50 PM   #2
mudshark
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this systems works really well:
http://www.chromotek.com/products/GFP-Trap.html

as for 'normal' ABs, unpurified polyclonal sera usually work best in my hands, monoclonals are usually less preferred.
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Old 01-28-2012, 07:40 AM   #3
osl
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Thank you mudshark. I'll try this system.
Have you been working with the GFP-Trap agarose or the magnetic ones? Looks like the agarose beads have a higher binding capacity.
Thanks again.
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Old 01-28-2012, 08:29 AM   #4
mudshark
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agarose only, no idea about the magnetic ones.
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Old 08-21-2012, 01:23 AM   #5
Buchi
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Default ChIP-Seq with GFP-Trap agarose beads

Quote:
Originally Posted by mudshark View Post
this systems works really well:
http://www.chromotek.com/products/GFP-Trap.html

as for 'normal' ABs, unpurified polyclonal sera usually work best in my hands, monoclonals are usually less preferred.

Hi,

I would also try the GFP-Trap for my experiments. Do you have a good working protocol?

Cheers
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Old 09-28-2012, 09:30 AM   #6
epigeneticfan
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Quote:
Originally Posted by osl View Post
Hi all,
Just wondering if anyone could recommend a good anti-GFP/YFP for ChIP-Seq.
I've tried quite a few (including ab290, JL-8) for ChIP-qPCR. I just found the polyclonal gives rather high background, and the monoclonal seems to have low efficiency (I'm working on Arabidopsis).
Also, for ChIP-Seq, is monoclonal Ab a better choice over polyclonal?
Thanks ahead.
os

There is no general rule for using monoclonal or polyclonal antibodies, but the main differences are the following:

The big advantage of the polyclonal is that it will recognize several epitopes on the same antigen. Which increase the change to recover enough material, especially for sequencing. But you have batch-to-batch variation. It might be that a monoclonal is “too specific”, it recognized only one epitope. And in some cases this epitope might be hidden (because of the fixation with formaldehyde leading to a different chromatin structure. All polyclonal antibodies are different so you will have to try by yourself.

The advantage of the monoclonal is that once you have a good one and it works you don’t have to worry about batch-to-batch variations.

Coming to the antibody: what we have in our catalogue is the following antibody:

Validated in IF, but not yet in ChIP.
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