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Anti-NCoR2 antibody [1212] (ab2780)

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2 questions for ab2780

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Question 1

Tuesday 17-May-2005

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM High background, low signal and non specific staining

SAMPLE HeLa cells

PRIMARY ANTIBODY NoCR ab2780, diluted 1/100 in PBS 10% FBS

ANTIBODY STORAGE CONDITIONS -20?

FIXATION OF SAMPLE Paraformaldehyde 4%, 10 minutes room temperature

PERMEABILIZATION STEP PBS-0.1 % Triton X-100

BLOCKING CONDITIONS PBS 10% FBS

SECONDARY ANTIBODY Cy5 anti-mouse, diluted 1/100 in PBS 10% FBS

ANSWER:

  

I'm sorry to hear you are having a problem with ab2780.

We have not had any complaints regarding this antibody, so I believe that I can give you some protocol tips to get the antibody working. I believe that the problem may be in the blocking step using 10% FBS, or a different fixation may also help. This antibody has been tested in immunohistochemistry (Frozen sections) in HeLa cells and I am assuming this is the application that you are using. I would like to suggest the following modifications to your protocol: 1) block using 10% normal goat serum 1% BSA in TBS for 2 hrs at room temp, 2) incubate in primary antibody at 1/50 in 1% NGS 1% BSA O/N at 4C, 3) incubate in secondary antibody in 1% NGS 1% BSA for 1 hr at room temp. Additionally, you could try fixing using ice cold acetone or methanol for 10 minutes instead of PFA, if the above suggestions do not work.

Please let me know if this helps and do not hesitate to contact us for further advice,

Question 2

Thursday 16-September-2004

Does this antibody recognize NCoR1 as well as NCoR2?

ANSWER:

  

This antibody has not been tested for cross-reactivity with NCoR antigens. We have an NCoR antibody, ab3482. If you have any further questions, please contact us again.

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