joan@uncmed.med.unc.edu (08/16/90)
If there is anyone out there in netland who has worked with membrane proteins (esp. in assay situations), PLEASE, I NEED HELP AND LOTS OF ADVICE!! Thanks, Joan Shields UNC-CH Dept. of Anesthesiology "Will you miss me, will you miss me, miss me when I'm gone?"
joan@ocean.med.unc.edu (Joan Shields) (08/24/90)
In article <840@beguine.UUCP> joan@uncmed.med.unc.edu () writes: >If there is anyone out there in netland who has worked with membrane >proteins (esp. in assay situations), PLEASE, I NEED HELP AND LOTS OF >ADVICE!! 've gotten lots of replies to my query and I would like to thank all who have answered my plea for help. I guess I should have been more detailed about what I am interested in. I am working with c-fos proteins in rat brains. I have been using an ELISA assay, which I have had little trouble with, however I have been having problems finding a detergent that will either not interfere with the ELISA or that is easily removed. References, and of course any hints and advice, will be happily received. Thanks again. Joan Shields UNC-CH "There's rumors in the loading bay there's anger in the town Somebody blew the whistle and the walls came down"
v151luy2@ubvmsa.cc.buffalo.edu (Scott D McGinnis) (09/10/90)
In article <915@beguine.UUCP>, joan@ocean.med.unc.edu (Joan Shields) writes... >In article <840@beguine.UUCP> joan@uncmed.med.unc.edu () writes: >>If there is anyone out there in netland who has worked with membrane >>proteins (esp. in assay situations), PLEASE, I NEED HELP AND LOTS OF >>ADVICE!! > Actually, I tried to E-Mail you but for some reason you are out of my jurisdiction. I have alot of experience w/ ELISAs and have worked purifying the Rhodopsin/GTP-ase/PDE-ase sytem *functionally* What I don't understand is the nature of you ELISA problems? Be exact I can understand any tech. jargon. As for the C-fos system which I am unfamiliar, can you tell me weither it is the G-protein in the cascade or does it play another role? Is the detergent necessary to purify it, or is the detergent in the ELISA wash buffer (your wording is unclear)? Do you need help purifying other enzymes such as the membrane protein in your system which activates the G-protein and the Adenylate Cyclase (if that's what it is)? And I am interested in *how* you intend to assay this system: for instance, the formation of cAMP ect.? Finally, does the system need to be reconstituted into mem- brane vesicles in order to be functional (a potent- ial pain in the rump right here)? I know that this is a lengthy cross-exam. but I can handle any technical jargon I am quite familiar with both of these systems as I've said. You may want to E-mail me direct so as not to clutter up the newsgroup. Scott V151LUY2@ubvmsa