I'm having some trouble trying to stain for endothelial cell in rat brain tissues. My positive control works for CD31, but my samples are not coming up. I'm wondering if it is my tissue preparation that is the problem.
If anyone has a protocol for the tissue preparation and staining that would be greatly appreciated!
Hallo Shi, if your positiv control is treated in the same way as your brain tissue and you have a positiv result in your control material and nothing in your brain tissue, you have to check weather the control tissue is also from rats. If not the rusult shows you that this CD31 antibody is not suitable for rat tissue. If your control tissue is from the same species like the brain tissue, in that case the result shows you that the protein CD31 is not detactable in brain tissue. That is exactly the reason why you are running positiv controls. Positiv controls show you that the immuno protocoll is woking even when you could recieve negativ results. In that case you have to look for another CD31-AB wich is suitable for your kind of tissue.
Hi Ute,
Thank you for your reply. Both my positive control and control tissue are from rat tissue. I read online that certain methods of extracting and fixing the brain can affect staining of blood vessels, and was wondering if anyone knew the best way to prep brain samples before staining.
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