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Anti-STAT3 (phospho Y705) antibody (ab30646)

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1 question for ab30646

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Question 1

Tuesday 27-September-2011

I have a question regarding the treatment of tissue prior to applying the primary antibody. Do you recommend microwave? Enzymatic?

The tissue I have in mind is human salivary gland.

I'm planning to use ab6672, ab59389, ab39636, ab32101, ab31370 and ab30646. Different recommendations?

ANSWER:

  

Thank you for contacting us.

The treatment of PFA-fixed, paraffin embedded sections (IHC-P) before incubation with priamry antibody would include antigen retrieval, blocking of endogenous peroxidases (if HRP is being used for detection), and blocking of non-specific proteins. Here are the protocol details for the respective antibodies:

ab30646, ab31370, ab39636, ab59389:

1. antigen retrieval: 10 mM citrate buffer (pH6.0) boil in pressure cooker

2. block with 3% H2O2 (in fresh methanol) for 15 minutes at room temperature

3. block with 3% BSA in TBS for 30 minutes

ab6672:

antigen retrieval: citrate buffer pH 6 and proteinase K digestion have been tried successfully as well as no antigen retrieval.

2. block with 3% hydrogen peroxide for 10 min

3. block with 1-10% Serum for up to 1 hour

ab32101:

1. antigen retrieval:10 mM Sodium Citrate Buffer, pH 6.0 in rice cooker

2. block with 3% hydrogen peroxide for 10 min

3. block with PBS + 10% serum

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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