Thursday, April 14, 2016
Week 9 and Protocols
This week I did a Fluorescent Immunohistochemistry. It's basically the same procedure except the secondary antibody is fluorescent, the primary antibody is a lot more concentrated, the tissues must be kept in the dark after the secondary is added, and a few other things. I made a mistake during the experiment. Generally, we can put two antibodies together but only if they are made in two different animals. I was targeting tau and amyloid proteins so I put both their antibodies in. After 2-3 hours I realized that both antibodies were made in mouse (so I wasn't supposed to put them in together because then the secondary could bind to either and it would not differentiate) so I washed the tissue as much as I could in hopes that the antibody hadn't blinded yet. Then I put only one antibody in for amyloid plaques. Hopefully the results are okay. I briefly looked at them under the microscope and it seemed fine.
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Kinza: How did the Fluorescent Immunohistochemistry stain work out? Were you able to successfully tag the plaques? I am eager to see the results!
ReplyDeleteIt was successful but because I put both antibodies by accident for a few hours, there is some tau staining too. I think this is actually a good thing because I can show some tau and amyloid together which is what I planned to do in the first place (but it was supposed to be two different colors). The results are also not as bright as they should have been but they are definitely very clear.
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