Antibodies for Immunofluorescence: What Do You Use?
Hey folks, I’m setting up a panel for immunofluorescence staining in mouse tissue, and I’m trying to decide which antibodies are the most reliable. I’ve had mixed results with some commercial ones—either they give off weak signals or the background is just too high to interpret. Anyone here got recommendations for antibodies that consistently perform well, especially for neural markers like GFAP or Iba1? Would really appreciate any pointers before I start wasting time (and money) again.
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I totally get your frustration—been there, done that. For GFAP, I had issues with signal fading using one of the standard suppliers, but then I switched to a rabbit polyclonal from Gentaur and it gave me beautifully clear astrocyte staining in frozen mouse brain sections. Iba1 was trickier—I went through three different brands before landing on a goat antibody that worked well in 30μm vibratome slices. If you haven’t checked out Gentaur yet, they’ve got a surprisingly decent catalog of validated antibodies, especially for neuroscience work. You might want to browse around here: what is chromatography — I found it useful because they categorize everything by application and host, which makes filtering way easier. Also, double-check if your secondary is getting cross-reactivity—once I switched to highly cross-adsorbed secondaries, my background dropped drastically.