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BATCH NUMBER 134780 ORDER NUMBER 111474 DESCRIPTION OF THE PROBLEM No staining visible on positive control of tonsil. SAMPLE Tonsil, Placental Tissue PRIMARY ANTIBODY ab 15497 (Rabbit Polyclonal to PCNA) 1:200 diltution incubated at RT for 10 min. Washed with PBS with Tween DETECTION METHOD Confocal fluoresence microscope POSITIVE AND NEGATIVE CONTROLS USED Tonsil - Positive control ANTIBODY STORAGE CONDITIONS 4oC FIXATION OF SAMPLE Formalin ANTIGEN RETRIEVAL Enzymatic method using Proteinase K - After de-waxing, incubated in solutions of Proteinase K 20ug/ml for 20 min at room temperature. PERMEABILIZATION STEP NA BLOCKING CONDITIONS NA SECONDARY ANTIBODY Alexa 647 from Molecular Probes A-21246 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? None
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ANSWER: |
Thank you for your enquiry. I hope this information helps, please do not hesitate to contact us if you need any more advice or information, I'm sorry to hear you are having a problem with ab15497. I would like to suggest the following modifications to your protocol: The datasheet states the following: No special pretreatment is required for the immunohistochemical staining of formalin/paraffin tissues. It is possible that antigen retrieval destroys the epitope. Have you tried staining without performing antigen retrieval? Please let me know if this helps and do not hesitate to contact us for further advice. |
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