Thermo Scientific Pierce
Donkey anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 555 conjugate from Thermo Scientific Pierce Antibodies
A-31572
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Secondary antibodies from Thermo Fisher Scientific are great for doing immunofluorescence. We work with primary cells (airway epithelial cells) in our lab and can use several different donkey alexas with different fluorophore conjugations in a single staining without any issue. A dilution of 1:2000 for 30 - 60 minutes works great with most of them.
Immunofluorescence
Normal human bronchial epithelial cells
Usually overnight at 4 degrees and dilution depends on primary Ab in 3% BSA
3% BSA
1:2000 ; 30-60 minutes at room temperature in 3% BSA in the dark
DAPI in PBS 5' RT
Confocal microscopy
Can use up to 3 different donkey AF (3 different species and 3 different fluorophores) without any issue in background or cross reactivity.
None
Easy to optimize, lasts forever
When it gets old, starts aggregating
Great product!