Anti-NCOR2 antibody - ChIP Grade (ab24551)

What I wanted to know as far as positive control was something to the equivalent of "did they use rabbit whole serum or rabbit IgG as a control in the IP portion of the assay?" One has to use a positive control in this assay and if you look at those publications they show IgG in their figures. I am shocked that you don't test these internally in your ChIP system and rely solely on a customer's results - no internal standard testing is almost unbelievable.

ANSWER:

Thank you for your enquiry.

We qualify our "ChIP grade" antibodies in three ways:

Through our; 1. In our in-house ChIP testing program, 2. In a scientific publication or 3. by a customer review.

On this occasion this antibody was sourced from the authors. We have every confidence in the behaviour of this antibody by ChIP.

We make every effort to test as many of our antibodies in house as possible. However, this is not always possible given the cell systems employed to detect the binding of chromatin associated proteins. We only qualify our antibodies as being suitable for chromatin immunoprecipitation following a customers review if we are aware of the protocol used and the necessary optimisation for this antibody. We also monitor the status of our ChIP grade antibodies very closely. Should a ChIP grade antibody be shown to no longer demonstrate the capacity to immunoprecipitate cross linked chromatin we remove ChIP as an application.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

What was used as a positive control in testing these antibodies in ChIP?

ANSWER:

Thank you for your enquiry.

We qualify our antibodies as "ChIP grade" status in three ways 1. in our in-house ChIP testing program, 2. in a scientific publication and 3. by a customer (Abreview).

Rabbit polyclonal to NCoR2, ab24551 was qualified as being capable of ChIP by its use in the following publication:

Yoon HG et al. Reading and function of a histone code involved in targeting corepressor complexes for repression. Mol Cell Biol 25:324-35 (2005). PubMed: 15601853

In this publication they demonstrated the association of NCoR with the deiodinase 1 (D1) promoter (thyroid hormone response element (TRE)) in a HeLa a2 cell line which constitutively expresses a FLAG-tagged human TRa. They did not use a positive control as such. However, if this does not present itself as a suitable control any thyroid hormone controlled gene that is reactivated with Hdac inhibitors or has been shown to have association with NCoR are good places to start.

ab24552 was also qualified by its presence in the same publication. It was used and confirmed by the same authors.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.