Re: [Histonet] Immunogold question

From:Robert Chiovetti

Hi Danielle,Just a guess here, but the higher pH may help reduce nonspecific background staining.  I worked in an EM lab that occasionally played with pH and salt concentrations for immunogold labeling when it was necessary.  I know for sure that high salt washes after the primary and secondary Ab's, followed by a return to normal TBS, does a super job of getting rid of background.  I'm not so certain about the pH.  I only used it a couple of times and I really couldn't tell much difference...Bob =0ARobert (Bob) Chiovetti, Ph.D.Southwest Precision InstrumentsArizona's Microscopy Resource132 North Elster DriveTucson, AZ 85710-3212Tel./Fax 520-546-4986Member, Arizona Small Business Association(www.asba.com)----- Original Message ----From: Danielle Crippen To: histonet@lists.utsouthwestern.eduSent: Monday, January 29, 2007 10:27:46 AMSubject: [Histonet] Immunogold question=0ADear EM experts,I've been using the following protocol for immunogold labeling for the past couple years with good success.  This morning one of our users has inquired as to why the secondary is incubated at a higher pH than the primary.  I do not know the answer...do any of you???=0A_______________________________________________Histonet mailing listHistonet@lists.utsouthwestern.eduhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet ____________________________________________________________________________________Want to start your own business?Learn how on Yahoo! Small Business.http://smallbusiness.yahoo.com/r-index
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