Lately I’ve been tweaking an IF setup and noticed the signal behaved very differently depending on the secondary antibody. When does using an anti-IgM secondary really outperform a pan-immunoglobulin approach for cleaner amplification? I once spent a whole weekend chasing background haze, and that experience still makes me second-guess my choices.
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Back when I was working with some early B-cell samples, I leaned toward anti-IgM secondaries because the readout felt more focused to me, especially when the expression was low. With pan-Ig detection, I sometimes felt like everything lit up at once, which looked impressive but got messy fast. Reading up on the lgm full form helped me put words to what I was already seeing at the bench. That said, it’s still a bit of a judgment call, and I don’t think there’s a universal “better” option.