what does in mean by 2x buffer solution when preparing a protien sample for SDS PAGE?

Question

I have a 5 microliters of protien sample and I need to prepare a 15microliters for and SDS PAGE with2x buffer solutiona d dH20

Answer 1

2x means that the concentration of the components in that solution is double their concentration in the final solution. So you need to mix equal volumes of sample and buffer solution to have the right concentration.

In SDS PAGE you could easily just put 5 microliters of sample and 10 microliters of loading dye (your 2x buffer solution) and there won't be a difference.

If you really want to be exact you need half of your final volume being the 2x solution and half sample. Thus for 15 microliters total you mix 7.5 microliters 2x solution, your 5 microliter sample and 2.5 microliters dH2O

Answer 2

The denaturing buffer used in sample preparation for SDS PAGE (sodium dodecyl sulfate - polyacrylamide gel electrophoresis).

5 microliters of protien sample + 10 microliters of 2X buffer will give 15microliters to use on the gel.

Answer 3

2x Sds Loading Buffer

Answer 4

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Answer 5

Hey, you have not mentioned the pH of your stacking and separating gels. We did SDS PAGE from an educational kit so we only had to dilute the sample loading buffer. However, pH of stacking gel was 6.8 pH of separating gel was 8.8 It might not be the same for your experiment, but just thougt I'll let you know.

Answer 6

concentrated
2 x normal recipe
if buffer solution is 10% NaCl then 2X would be 20% NaCl.