Role of glycine based blocking in Immunofluorescence - (Feb/06/2008 )

Hi everybody. I've got a very simple question: I've been told that glycine might help decreasing the staining background in immunofluorescence microscopy. Do you know if it actually works, and, if so, how does it work?

thx very much

dont know about glycine but you could try cold fish skin gelatin (sounds bizarre but i'm assured it works) - combine it with triton x in a little pbs with bsa and the serum of your choice
i'm trying it myself next week hopefully
if the glycine works let me know how it goes

dom

never heard of glycine in immunofluorescence. but if it works please do inform us.

ASAP, glycine can be used in immunofluorescence assay as quenching agent to be used after fixation (in the case of PFA fixation). Indeed, glycine reacts with the excess of formaldehyde, thus stopping the fixation reaction. To my experience, simple and abundant PBS washing after fixation is sufficient to remove unreacted aldehyde.

I also do treat my paraffin embedded with 100mM glycine in PBS for 45 min.
That's because the step is on my protocol that I got.
But have no idea what it does, besides, I dont see much difference anyway
or maybe I am doing something wrong.

also I treat slides with 50mM NH4CL in PBS for 15 min, which I assume to lyse RBC, but it also has very few effect on my lung sample.

do you guys have any other methods to remove RBC??
(BTW I tried Methanol:Acetic acid, it didnt do anything)