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Colocalization - Fluorescence Histological Double Labeling (Nov/04/2004 )

I'm trying to colocalize Tyrosine Hydroxylase and the GDNF receptor (GFRa1). Tyrosine Hydroxylase labeling (Rhodamine conjugated secondary antibody) works OK but I'm having problems with FICT conjugated secondary antibodies as I get to much background and poor cell labeling. I've tryied to dilute secondary antibodies but with little labelling improvement. Hope someone can help with this, Josť W.

-boina-

a) you can reduce your background by treating your samples with 1 mg/ml sodium borate (in PBS) for 10 min after you've permeabilised and before you incubate with primary.

cool.gif Also, try to filter your antibodies through a 0.2 micron filters (micro filters for syringes are available). Both your primary and your secondary. This way you'll avoid big clumps on your samples.

good luck,...

-Sprag-