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  • Help with dilutions

    Hi,
    I'm preparing a protocol in which I have to make several dilutions of a target protein. The stock acquired has a volume of 100μL in a concentration of 100 μg/mL, and a molecular weight of 5807.57. I am aiming at having 5nM, 10nM, 1μM, and 5μM final concentrations in a final mix solution volume of 10 μL for each. Can someone guide me?
    Thank you!

  • #2
    Originally posted by Joao View Post
    Hi,
    I'm preparing a protocol in which I have to make several dilutions of a target protein. The stock acquired has a volume of 100μL in a concentration of 100 μg/mL, and a molecular weight of 5807.57. I am aiming at having 5nM, 10nM, 1μM, and 5μM final concentrations in a final mix solution volume of 10 μL for each. Can someone guide me?
    Thank you!
    I think the first step is to express your stock solution in molarity. "M" is shorthand for moles per liter, so you have
    100 痢/mL x 1 mole/5807.57 g x 1000 mL/L = 17.22 痠ol/L (or 然). At this point you can use C1 * V1 = C2 * V2

    I would make up more than 10 無 of the 5 然 solution and use that for further dilution.

    If you take 5 無 of the stock and dilute it to 17.22 無 final volume, you will then a new stock of 5 然. Reserve 10 無 of it for that requirement. Some of the rest will be diluted further for the other samples.

    For the 1 然 sample, take 2 無 of the 5 然 "new stock" and dilute to 10 無

    The other two are very dilute and may require two step dilution, but 5 然 is 5000 nM. so dilution to 10 nM requires 500:1 dilution. Take 1 無 of the 5 然 solution and dilute to final volume of 500 無. You now have lots of extra 10 nM solution, reserve a 10 無 sample. Then take 2 無 of it and dilute to 10無 for the 1 nM solution.

    Your solutions are so dilute volumes are probably additive, but they are not for strong solutions, so "best practice" is always to dilute to a final volume, not to add a specified volume of water or solvent.

    Comment


    • #3
      Originally posted by JohnS View Post

      I think the first step is to express your stock solution in molarity. "M" is shorthand for moles per liter, so you have
      100 痢/mL x 1 mole/5807.57 g x 1000 mL/L = 17.22 痠ol/L (or 然). At this point you can use C1 * V1 = C2 * V2

      I would make up more than 10 無 of the 5 然 solution and use that for further dilution.

      If you take 5 無 of the stock and dilute it to 17.22 無 final volume, you will then a new stock of 5 然. Reserve 10 無 of it for that requirement. Some of the rest will be diluted further for the other samples.

      For the 1 然 sample, take 2 無 of the 5 然 "new stock" and dilute to 10 無

      The other two are very dilute and may require two step dilution, but 5 然 is 5000 nM. so dilution to 10 nM requires 500:1 dilution. Take 1 無 of the 5 然 solution and dilute to final volume of 500 無. You now have lots of extra 10 nM solution, reserve a 10 無 sample. Then take 2 無 of it and dilute to 10無 for the 1 nM solution.

      Your solutions are so dilute volumes are probably additive, but they are not for strong solutions, so "best practice" is always to dilute to a final volume, not to add a specified volume of water or solvent.
      Thank you so much! That's exactly what I needed

      Comment


      • #4
        Helpful tips...thanks

        Comment

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