I have used a Tyrosine hydroxlase antibody from Eugene Tech International
INC. at 1:500 for 48 hrs at 4-8degrees C, we tried RT and shorter
incubation times and were not happy with the results. It was an Alk/phos
protocol with Fast Red Chromogen.  It has been awhile but as I remember the
results were pretty consistent.
Gina Sennello
Gilead Sciences




                                                                                            
                    Anila Syed                                                              
                    <syedab@total        To:     'histonet' <Histonet@pathology.swmed.edu>  
                    ise.co.uk>           cc:                                                
                                         Subject:     Tyrosine Hydroxylase                  
                    05/01/01                                                                
                    08:48 AM                                                                
                                                                                            
                                                                                            




Dear All,

Sorry if this has been asked before. I have a protocol for Tyrosine
hydroxylase which I am using to stain 50um brain stem sections. It has a 4
day incubation of primary on it at 4 deg C and I was wondering whether
anyone has had any experience using this ab. My question is, can I do a
shorter incubation at room temp and get the same results?

A secondary question I have is how reliable is immunostaining for tissue in
which you want to count and measure cells? Does one get consistent results
with a given ab and a given protocol? i.e. if one used the same protocol on
the same tissue section, would the same cells always be stained and to the
same extent?

Many Thanks in Advance,

Kind Regards,


Dr. Anila Syed