Chemical Forums
Specialty Chemistry Forums => Biochemistry and Chemical Biology Forum => Topic started by: payata on December 17, 2019, 08:21:10 AM
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Hi Group!
I dealing with a dilution of antibody: my final volume is: 10 ml, and dilution must be done 20000x
I think but I am not quite sure I need to do it stepwise, can someday negative, which examples, how and from what volume/dilution to start,
Thank you!
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Suppose that you did not do this stepwise. What volume would you need for a 20,000-fold dilution?
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10 ml
I know formula DF= Vf / Vi = but 0,0005 ml, which is 0,5 ul in 9,9995 ml volume? I think this is not possible
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It would be very difficult for me to do this accurately, but it depends on what equipment one has. If we rule out doing the dilution in one step, then it would have to be done serially. You have some choices about how you could do this, but as long as the two dilution factors multiply to 20,000, you should be OK.
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OK, but can anyone show me an example, how to make it stepwise?
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If I were preparing the antibody dilution, I would just add 0.5µL of the antibody into 10mL of the appropriate buffer. For most applications (e.g. Western blotting) the concentration does not have to be very precise, so there is no need to take steps to make a more accurate dilution.
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It is for ELISA, and one of the most frequent questions is to make 20000x dilution, 25000x etc When I asked Student who had this test before, they said that Teacher wants us to make stepwise, because nobody had this question correct until now, so I am a bit confused
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According to the Forum Rules (see red link above), you must show us your attempt before we can help you.