Question about fixation for immunocytostaining - (Jan/09/2006 )

I'm asking this question because I'm kinda lazy today. Can you stain cells with a primary antibody, formalin fix it, and then procede with a secondary antibody later? or incubate with a biotinylated primary, fix, and then add steptavidin conjugate later??

you fix after incubating with the primary????

we fix, permeabilize, and block; then o/n with primary antibody at 4 C

makes a good spot to stop for the day

I did not know you could fix after adding the primary

I'm staining surface protein on live cells for flow. primary is added to live cells for 1hour @ room temp then washed. we then add secondary etc.. then fix. What I wanted to know is ... if I fix following the primary step, will the secondary recognize the fixed primary. I've never done this and am hesitant to try, but it would save me a couple of hours tonight. My ussual motto is to due things right the first time, but this little cheat will get me home for dinner .

you fix after incubating with the primary????

we fix, permeabilize, and block; then o/n with primary antibody at 4 C

makes a good spot to stop for the day

I did not know you could fix after adding the primary