I'd like to preface that I haven't done any Alexaflour testing in just over 10 years. The simplest response may prefer using a conjugated secondary if you're planning to do multiplex or multicolor analysis.
If I was limited in commercially available options for primaries needed for the test sample, then i would prefer a conjugated secondary as it loosens up testing requirements for testing. Another benefit is that it may increase specific testing and allow positive staining to appear more visibly present than a directly conjugated antibody. There are other benefits for a conjugated secondary or tertiary antibody, but it requires users to know how to troubleshoot the stain. Direct conjugation is easier to use in many circumstances. I would communicate to your friend to try both for the study and determine as a team of internal collaborators on which method is easiest and adequate enough for performing the method testing. I hope this helps. Eddie NIH Bone Marrow Service RND clinical & anatomic pathology specialist > On Oct 19, 2023, at 9:22 AM, Charles Riley <cri...@udel.edu> wrote: > > Why would one decide to use a primary antibody along with a secondary > antibody rather than a primary antibody conjugated with the secondary? > > Example. I have a researcher who wants to do CD11C staining with > Alexafluor488 > > Is it better to buy and use a primary antibody CD11C conjugated with > Alexafluor488 or to do the CD11C primary and a Rabbit anti-rat (H_L) IgG > antibody secondary? > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
