EveC

Hello, thanks for your reply. Yes, of course, you have to be precise in science and agree with a misunderstanding when asking people. Other techniques, for example, in molecular biology, will need all the precision, and this type of rounding affects the result. The immunofluorescence worked to show the presence of a protein in the tissue region. For now, I have not done a quantification with the IF. However, in the future, it is better to be careful with this rounded. Thank you and have a nice weekend!

Hello everyone, and thank you very much for your very helpful responses. The confusion came mainly from the way some people say: "a dilution of 1: 100 so add 3ul in 300ul of buffer", which is not accurate, as you mention. However, in some biological techniques, the precision also depends on the method and objective, as you also precise. In particular in histology (I am working for now with histology techniques). This is the case in immunohistochemistry. For an immunofluorescence assay, can this type of rounding be done? Thank you very much and have a nice day.

Hello, I would have a question about dilutions in immuohistochemistry. Maybe could help me? The desired dilution is 1: 100. The first protocol that I used said to add 10ul in 1000ul buffer. I considered this too much antibody, but, If I am right, it is still correct since 1000/10=100 dilution factor. After that, a new protocol was available, and this last protocol said to do 1:100 dilution and prepare 500 ul of primary antibody solution. So I prepared 5ul antibody and 500 ul buffer (since 500/5 = 100, dilution factor). However, I see that some people consider that it has to be 5ul and 495ul of buffer to reach the final volume of 500 (or in the first case 10ul antibody and 990ul buffer). Which of these ways to do is valid? Is this correct to add 5ul of Antibody in 500ul of buffer to have a dilution of 1:100? I am confused concerning total volume. Thank you very much for your time and help. I look forward to hearing from you.