Lec 8

Question 1

Isolating proteins purpose

Answer 1

Proteins can be isolated from tissues, cells or organelles

Question 3

Groups

Answer 3

Experimental group

Positive/Negative control

Negative- event should not occur

Technical control- Allow you to quantify the results of all groups

Question 4

SDS PAGE

Answer 4

Separate protein by size by moving them toward a positive electrode

Done by coating all proteins with a negative charge because not all proteins are negative on their own

Next, hinder the protein movement with a fibrous gel

The proteins will move towards the electrode but are slowed down by the fibers

The smaller proteins will move faster through the gel

Question 5

SDS page

Lane

Answer 5

Because the proteins are trying to take the shortest route, they run in a lane

Question 6

SDS page

Band

Answer 6

Where the proteins end up

Lanes can have multiple bands

Question 7

Coomassie blue staining

Answer 7

Shows all proteins on a gel

Soaking gel with coomassie blue reveals proteins

Question 8

Technical control of SDS page

Answer 8

Molecular weight marker (lane)

Tells us the size of our band

Use proteins with known sizes

Question 9

Western blotting

Answer 9

Shows a specific protein on a gel

Each band shows many copies of one protein

Transfer the proteins onto a membrane (blotting)

All proteins can be seen on the membrane including YFP

Detected using antibody attached to an enzyme

Any antibodies that didnt stick are washed off, only leaving YFP

Detection reagents are added and when reacted with the enzyme, make a blue precipitate

Question 10

Loading control

Answer 10

Type of technical control

Antibodies against a protein that should be present in all samples

Question 11

Molecular weight markers work for

Answer 11

Western blots and Coomassie blue staining