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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? |
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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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Immunohistochemical analysis of paraffin-embedded breast carcinoma. Left: Using anti-STAT3 antibody(ab31370, diluted 1:100); Right: The same antibody preincubated with synthesized peptide.
All lanes : Anti-STAT3 antibody (ab31370) at 1/1000 dilution
Lane 1 : K562 cell lysate
Lane 2 : K562 cell lysate with synthetic non-phosphopeptide
Predicted band size : 88 kDa
Observed band size : 95 kDa (why is the actual band size different from the predicted?)
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