L11 - How are Proteins Studied with GELS?

0.0(0)
Studied by 0 people
call kaiCall Kai
learnLearn
examPractice Test
spaced repetitionSpaced Repetition
heart puzzleMatch
flashcardsFlashcards
GameKnowt Play
Card Sorting

1/7

encourage image

There's no tags or description

Looks like no tags are added yet.

Last updated 4:52 AM on 5/24/26
Name
Mastery
Learn
Test
Matching
Spaced
Call with Kai

No analytics yet

Send a link to your students to track their progress

8 Terms

1
New cards
<ol><li><p>Isolating proteins</p></li></ol><p></p>

  1. Isolating proteins

  • proteins can be isolated from tissues, cells, or organelles → get total proteins

via : ToPI-blood = a method of isolating proteins from blood cells

→ then can use mass spec, western blotting, SDS-PAGE…

2
New cards

What method should be used to isolate mitochondrial proteins?

a). whole cells → subcellular fractionation → isolate proteins → mito. proteins

b). whole cells → isolate proteins → subcellular fractionation → mito. proteins

procedure A, as procedure B will contain other proteins

<p>procedure A, as procedure B will contain other proteins </p>
3
New cards
<ol start="2"><li><p>SDS-PAGE</p></li></ol><p></p>

  1. SDS-PAGE

= separate proteins on a gel

  • SDS - gives proteins a (-) charge

  • polyacrylamide gel - separates proteins by size

<p>= separate proteins on a gel</p><ul><li><p>SDS - gives proteins a (-) charge</p></li><li><p>polyacrylamide gel - separates proteins by size</p></li></ul><p></p>
4
New cards
<p>SDS-PAGE : method</p>

SDS-PAGE : method

  1. coat proteins in SDS ( now have neg. charge)

  2. load gel

  3. run gel

  • each band represents many proteins of a the same size (large higher, smaller lower to bottom)

NEXT: coomassie blue staining OR western blotting

<ol><li><p>coat proteins in SDS ( now have neg. charge)</p></li><li><p>load gel</p></li><li><p>run gel</p></li></ol><ul><li><p>each band represents many proteins of a the same size (large higher, smaller lower to bottom)</p></li></ul><p></p><p>NEXT: coomassie blue staining <u>OR</u> western blotting</p>
5
New cards

Control groups in cell biology: 2 types

positive control group = event should occur

negative control group = event shouldNT occur

6
New cards

Technical Controls in cell biology : 2 types

Molecular weight markers = tells us the size of our band

→ used in: coomassie blue staining and/or western blot

Loading control = antibodies against a protein that should be present in all samples

  • allows us to confirm findings aren’t due to forgetting to load proteins into a lane

ex. Bip, GAPDH, Actin

→ used in: western blots

<p><u>Molecular weight markers</u> = tells us the size of our band</p><p>→ used in: coomassie blue staining and/or western blot</p><p></p><p><u>Loading control </u>= antibodies against a protein that should be present in all samples</p><ul><li><p>allows us to confirm findings aren’t due to forgetting to load proteins into a lane</p></li></ul><p>ex. Bip, GAPDH, Actin</p><p>→ used in: western blots</p>
7
New cards

Coomassie Blue Staining

= shows ALL proteins on a gel (makes them blue)

  • used for verification experiments (ex. to make sure a plasmid transfected into E. coli is making proteins)

  • uses molecular weight markers (technical control)

  • depiction in journals: show most of gel

<p>= shows ALL proteins on a gel (makes them blue)</p><ul><li><p>used for verification experiments (ex. to make sure a plasmid transfected into E. coli is making proteins)</p></li><li><p>uses molecular weight markers (technical control)</p></li><li><p>depiction in journals: show most of gel</p></li></ul><p></p>
8
New cards
<p>Total proteins from bacterial cells before (lane 2) and after (lane 3) the plasmid was added.</p><p></p><p>Is lane 2 a positive control group or a negative control group?</p>

Total proteins from bacterial cells before (lane 2) and after (lane 3) the plasmid was added.

Is lane 2 a positive control group or a negative control group?

lane 2 is a negative control group