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If your product does not perform as described on this datasheet, we will refund or replace your product...
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Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
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ANSWER: |
Thank you for your reply. |
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DESCRIPTION OF THE PROBLEM No staining SAMPLE Mouse brain (fixed frozen) PRIMARY ANTIBODY ab2303, diluted in dako diluent or 1% BSA/0.3% triton X/PBS, diluted 1:30, 1:50 or 1:100, incubated at room temperature for one hour or overnight at 4C or at room temperature. DETECTION METHOD fluorescence or DAB POSITIVE AND NEGATIVE CONTROLS USED Negative: no primary antibody Positive: brain sections from mice that underwent motor cortex lesion 1 or 5 days before perfusion. These sections show positive TUNEL staining. ANTIBODY STORAGE CONDITIONS Stored in small aliquots at -20C. Aliquots are thawed and stored at 4C as needed. FIXATION OF SAMPLE formalin (perfused then post-fixed overnight) ANTIGEN RETRIEVAL Boiling citrate buffer for 15 minutes PERMEABILIZATION STEP 30 minutes in 0.3% triton X (triton X is also included in the antibody dilution solution) BLOCKING CONDITIONS Dako blocking solution or ImageIT FX signal enhancer or 1% BSA/0.3% triton X in PBS SECONDARY ANTIBODY anti-rabbit Alexa 488 (1:500 in dako diluent or 1% BSA/0.3% triton X/PBS for one hour at RT) or anti-rabbit biotinylated IgG (1:300 in Dako diluent or 1% BSA/0.3% triton X/PBS for 30 minutes at RT) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? antigen retrieval, blocking, concentration of primary antibody, duration and temperature of primary antibody incubation, detection method |
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ANSWER: |
Thank you for contacting Abcam regarding ab2302. |
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DESCRIPTION OF THE PROBLEM No staining SAMPLE Mouse brain (fixed frozen) PRIMARY ANTIBODY activated caspase 3 (ab2302)1:30, 1:50, 1:100 diluted in either Dako diluent or 1% BSA/0.3% triton X in PBS incubated for 1 hour at room temperature, overnight at 4C or overnight at room temperature DETECTION METHOD Fluorescence or DAB POSITIVE AND NEGATIVE CONTROLS USED Negative control: sections incubated without primary antibody Positive control: sections from mouse brain having undergone motor cortex lesion 24 hours or 5 days earlier. These sections stain positively for TUNEL. ANTIBODY STORAGE CONDITIONS Frozen in small aliquots at -20C. Aliquots are thawed and stored at 4C as needed. FIXATION OF SAMPLE formalin (perfused and post-fixed in formalin overnight) ANTIGEN RETRIEVAL I have tried boiling citrate buffer (15 minutes) PERMEABILIZATION STEP 0.3% triton X in the blocking solution (30 minutes) and in the antibody dilution solution. BLOCKING CONDITIONS I have tried commercial blocking solution, 1% BSA plus triton X in PBS, and Image-IT FX signal enhancer (30 minutes each) SECONDARY ANTIBODY anti-rabbit 488 1:500 for one hour anti-rabbit biotinylated antibody 1:300 for 30 minutes diluted in diluent or 1% BSA/0.3% triton X in PBS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? antigen retrieval, blocking solution, concentration of primary antibody, duration and temperature of primary antibody incubation, detection method (DAB and fluorescence) |
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ANSWER: |
I'm sorry to hear you are having problems with this antibody. Thank you for taking the time to fill in the questionnaire, it is very useful for me to understand your protocol. Your staining procedures and troubleshooting are excellent. |
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I really appreciate your help and your antibody suggestion but it would be more convenient for me to have a different host animal than rabbit. If i could request a refund that would be great. |
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ANSWER: |
Thank you for your reply. |
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Thanks very much for your prompt response. |
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ANSWER: |
Thank you for your reply. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
