Dear all,
I plan to make ChIP-seq for a specific transcriptional factor (TF) in D. melanogaster.
In this context, we have developed an ectopic protein (TF) tag with Flag: TF-Flag
We can precipitate the chromatin with Flag antibody.
I've two genotypes available the wild-type- flies (no TF -lag) & the TF-Flag- flies.
So at this time, I've made several ChIP and validated the specificity by qPCR using different controls.
I tested four possible controls.
- Input in flies - in TF_Flag- flies
- No Antibody - in TF_Flag- flies
- Mouse - Antibody (Flag ab is made in mouse) - in TF_Flag- flies
- Flag ab in wild-type- flies
In all, I've a high enrichment for the target of my TF compared to these four controls.
Our results in qPCR showed the most aspecific binding was in wild-type- flies with Flag ab.
Do you think this is the best control?
I plan to make ChIP-seq for a specific transcriptional factor (TF) in D. melanogaster.
In this context, we have developed an ectopic protein (TF) tag with Flag: TF-Flag
We can precipitate the chromatin with Flag antibody.
I've two genotypes available the wild-type- flies (no TF -lag) & the TF-Flag- flies.
So at this time, I've made several ChIP and validated the specificity by qPCR using different controls.
I tested four possible controls.
- Input in flies - in TF_Flag- flies
- No Antibody - in TF_Flag- flies
- Mouse - Antibody (Flag ab is made in mouse) - in TF_Flag- flies
- Flag ab in wild-type- flies
In all, I've a high enrichment for the target of my TF compared to these four controls.
Our results in qPCR showed the most aspecific binding was in wild-type- flies with Flag ab.
Do you think this is the best control?
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