I am culturing undifferentitaed dental pulp cells and i want to stain for NLRP3 in the cell, i have the Millipore anti-NLRP3 antibody, it says on the site that they use 1:4000 antibody dilution for IHC but i want to do ICC?
Hi Banan,
I'd recommend doing a search on BenchSci (https://www.benchsci.com).
You can find out published data then check the M&M of paper for dilutions.
Please see attached screenshot.
I hope this helps!
Maurice
You should think of the manufacturer's suggested antibody dilution as a suggestion, not necessarily as something that is correct for your cells and your preparation. Its a reasonable starting point, but you'd benefit from trying a range of different dilutions. I know peole who recommend trying a 6 log unit range of dilutions in half log unit increments to identify the optimal range for the antibody in your specific preparation. We don't usually go to that extreme, but we bracket a couple log units to either side of the manufacturer's recommended dilution.
In terms of IHC vs. cell culture, our experience is that the optimal antibody dilutions for those two preparations are often pretty similar.
Dave Sherry
The basis for Dr. Sherry's comment is this. Take a look. One has a very difficult time picking the range de novo and thus I always recommend a 3 log range in half log units starting with 1:1000. That usually will pinpoint the best concentrations. It is important to use 48h incubations and stain properly.
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