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intestinal tumor as pos. crtl
no signal at all
4 um slides
AR: citrate buffer (heating plate)
block: 10% donkey serum 30min
ab16832: 5, 10 ug/ml
ab118519: 20 ug/ml
suggested: try other AR, use more Ab, try HRP-secondary + hematoxyline (to check if any signal can be obtained)
Asked on Apr 19 2012
Thank you for contacting us.
I am sorry to hear that you received so far no staining with the antibodies.
As we discussed by phone, I'd suggest trying the following:
1) try another antigen retrieval step (e.g. Tris/EDTA, pH 8)
2)use more priamry antibody
3) try using an HRP-conjugated secondary antibody and hematoxyline (for nuclear staining)- to check if any signal can be obtained at all, with the experimentalsetup you have.
Right now it is difficult to determine where the problem might stem from - there could be multiple causes. Thus, please let me know what results you obtained after trying these tips and we can discuss this further.
I wish you good luck and look forward to hear back from you.
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Answered on Apr 19 2012