Topic 15.5: SDS page

Question 1

what is SDS page used for

Answer 1

to separate proteins

Question 2

how does protein gel electrophoresis work

Answer 2

proteins are charged and will move towards anode or cathode if placed in electric field
proteins are separated base on size (mainly), isoelectric point, and pH ( if fully folded or native)

Question 3

what is required

Answer 3

gel - allows separation
buffer - maintain charge on protein samples
power supply - electric field
stain - identify

Question 4

what are the steps

Answer 4

1. protein samples into well
2. resevoir for buffer
3. small molecules = faster, bigger = slower

Question 5

what is sds

Answer 5

detergent which breaks non covalent interactions (no longer tertiary/secondary structure) = linear polypeptide chain and given fixed amount of negative charge

Question 6

why is a reducing agent used

Answer 6

reducing disulfide bonds

Question 7

what can it be used to find

Answer 7

determine molecular size by using molecular size of known proteins around it to estimate

Question 8

what can it be used to find

Answer 8

determine molecular size by using molecular size of known proteins around it to estimate
plot out molecular weight by distance migrated