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Product # ab14184 substance P antibody I am having a difficult time with this antibody for immunohistochemistry. Fix samples in 4% paraformaldehyde overnight. 1. Paraffin sections 2. Citrate unmasking PBS wash 3. Block 2-5% goat serum in PBS-1hr room temp 4. 1:1000-1:500 SubP antibody 4*C overnight PBS wash 5. 1:100 goat anti-mouse Alexa 488 or Texas red-2hr room temp PBS wash Nothing but background seen on rabbit sciatic nerve, rat tendon, testis, skin, heart, muscle and lung. All negative controls (no primary) tissues, especially rabbit nerve, have fluorescent areas within the tissues. Have no indication that this antibody is working in any of my tissues.
Asked on Jun 19 2013
We have seen that enzymatic antigen retrieval is very successful when using this product on tissue in IHC-P and would recommend this. Have you attempted use of this type of antigen retrieval? I have added a link to Abcam's antigen retrieval guide for your review below. Antigen retrieval guide: https://www.abcam.com/index.html?pageconfig=resource&rid=11488
Keith BAbcam Scientific Support
Answered on Jun 19 2013