You have been given a protein sample with an initial concentration of 125 microg

Question

You have been given a protein sample with an initial concentration of 125 micrograms/microliter. You will analyze this solution using SDS-PAGE, loading the gel with 20 micro grams and 30 micrograms of protein from your sample.

A) In the space below, clearly outline the dilutions(s) that you will perform (applying the dilution we discussed in lab).

B) What volume will you need to load for the 20 µg and 30 µg samples? Keep in mind, your concentration volume to load in the gel must be between 5 µL and 25 µL. Show your work.

Volume for 20 µg load:_____________________   30 µg load: _________________

Explanation / Answer

Stock concentration = 125 ug/uL

Dilution-I: Take 1 uL of the stock and make it up to 10 uL with the diluent.

Dilution-II: Take 1 uL of the dilution-I and make it up to 10 uL with the diluent.

Concentration of the Dilution-II = 1.25 ug/uL

So,

For 20 ug,

1uL ----> 1.25 ug

? uL ---> 20 ug

= 20/1.25

= 16 uL

For 30 ug,

1uL ----> 1.25 ug

? uL ---> 30 ug

= 30/1.25

= 24 uL

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