From a protein assay, you determine that your sample of purfied protein contains

Question

From a protein assay, you determine that your sample of purfied protein contains 2.5 mg protein/mL. you need to prepare a sample for sds-page. the final volume of the sample for sds-page should be 30uL and the final concentration of the sample buffer in the sds page sample is 1x. you would like to load a total of 5 ug of your purified protein on the gel. determine the volume of the purfied protein, volume 6x sample buffer, and volume water needed to preper a sample for sds-page ( recall final voulme should 30 uL) You must show your work for full credit.

No you do not need more information to solve. Please use units all time. EXPLAIN why each step was done. THank you

Explanation / Answer

Conc. of purified protein sample = 2.5 mg/mL = 2.5 ug/uL

Since we need 5 ug protein , so we will be taking 2 uL of this purified protein sample.

Total allowed volume = 30 uL

Remaining vol = 30 - 2 = 28 uL

The buffer needs to be diluted from 6x to 1x.

Since final volume has to be 30 uL , so for producing a 6 times dilution, we will take 5 uL of buffer. This will produe the 6 time dilution because this 5 uL volume is diluted to 30 uL.

So remaining vol = 28 - 5 = 23 uL

This much water has to be used.

So final answer is :

2 uL of purified protein sample + 5 uL of sample buffer + 23 uL of water = 30 uL final volume

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