Hi everyone,
I cut my first frozen sections, human tonsil, and I have stored them at -70. When I take them out to use, should I bring them to -20C and fix them right away? Or should I bring them to room temp and then pap pen/wash/block/wash them then fix them before adding my primary anti-body. My original plan was/is to bring them to room temp, pap pen them and then wash/block/wash fix. I am asking for your advice as my boss is away for two weeks and the protocol he left me with is a bit vague and requires some modifications to fit the research that we are doing. I was just wondering if there is a compelling reason to keep the sections at/near -20 after sectioning up through the fixation before starting the Immunohistochemistry. Also, we are using BSA as one of the components in our blocking solution as well as a component in our wash solution. Is there a danger in using lower/higher quality BSA? I'm a little worried that our BSA might contribute to increased background if its quality is not high enough. The BSA we may use is A-8327 Sigma, and I am thinking this might be better A3294, or one of these (A9647,A7906,A6793) This is a new job for me, and its been a long while since I have done any Immunohistochemistries.so I am going over the reagents we have on hand while I refresh my memory on immunohistochemistry and sectioning techniques. Thanks for all your support Patrick Patrick Lewis Research Associate II-Bench| Infections and Prematurity Seattle Children's Research Institute 206-884-1115 OFFICE 000-000-0000 PAGER 000-000-0000 CELL 206-884-7311 FAX patrick.le...@seattlechildrens.org OFFICE 1900 9th Avenue Seattle, WA 98101 MAIL M/S C9S-8, Seattle, WA 98101 WWW seattlechildrens.org <http://seattlechildrens.org/> CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet