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How do you decide the right dilution for your primary antibodies? I see so many different recommendations, and sometimes the datasheet suggestions don’t work at all. Is there a good rule of thumb, or do you just experiment until it looks right?
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Oh, the dilution game… I’ve wasted so many membranes because I was too confident in datasheet ratios. These days, I make a quick checkerboard dilution test before starting a new batch. Takes a bit more time upfront, but saves days of repeating experiments later. Also, using fresh buffer instead of old stock helps more than you’d expect — I learned that the hard way.
Honestly, finding the right antibody dilution is like trial and error at first. Even if the datasheet says 1:1000, your system might need 1:500 or 1:2000 depending on your secondary and detection method. I started keeping a dilution log for each antibody and noted what worked best. Gentaur’s product pages https://gentaur.co.uk/antibodies/ often include verified dilution recommendations from actual lab users, which helps narrow it down. My advice: start with the suggested range but always optimize for your own setup. It might seem tedious, but it saves time in the long run.