Protein Fractionation

0.0(0)
studied byStudied by 0 people
learnLearn
examPractice Test
spaced repetitionSpaced Repetition
heart puzzleMatch
flashcardsFlashcards
Card Sorting

1/9

encourage image

There's no tags or description

Looks like no tags are added yet.

Study Analytics
Name
Mastery
Learn
Test
Matching
Spaced

No study sessions yet.

10 Terms

1
New cards

Protein Fractionation

Separating proteins based on physical or chemical properties

2
New cards

How were proteins fractionated the PF lab?

Acid Precipitation and Salting Out

3
New cards

Acid Precipitation

Separation based on pH.

Precipitate unwanted proteins by passing through their isoelectric point

4
New cards

Salting out

Separation based on ionic strength

Dependent on hydrophobicity - proteins with more hydrophobic regions will precipitate first

Done at 4°C → decrease risk of inactivation

5
New cards

What are common issues that could occur during protein fractionation, and how do they affect the results?

  1. No or small pellet formation

  2. Low absorbance readings in CB assay

  3. Cloudy supernatant after centrifugation

  4. Overlap between fractions

  5. No clear trend in protein concentration across fractions

6
New cards

Why do we fractionate proteins?

To isolate proteins of interest based on specific chemical or physical properties

Increase accuracy for analysis such as electrophoresis

7
New cards

Reagents used in the PF lab

  1. Tissue Extract

  2. Protein Buffer

  3. Acid Precipitation Buffer

  4. Ammonium Sulfate Salt Solution

  5. CB Protein Assay Reagent

  6. Protein Standard

8
New cards

Aims of the PF Lab

Learn protein purification strategies

Lean protein fractionation principles

Protein fractionation by changing pH/salting out

9
New cards

PF Methods

  1. Precipitate the proteins with an unwanted pH through acid precipitation

  2. Precipitate the proteins that are more hydrophobic through salting out at 3 levels of ammonium sulfate concentration

10
New cards

Protein Fractionation principles learned

  1. Isoelectric Precipitation

  2. Salting Out