Lecture 6 - Extraction and Analysis of Proteins From Cells 1 Things to Learn in This Week's Lab 1 How to extract proteins from cells 2 How to make

Lecture 6 - Extraction and Analysis of Proteins From Cells...

This preview shows page 1 - 9 out of 23 pages.

Electrophoresis and Electroblotting of Proteins 1 10/11/10 1 Extraction and Analysis of Proteins From Cells 10/11/10 2 Things to Learn in This Week’s Lab 1. How to extract proteins from cells. 2. How to make parallel dilutions from a stock solution. 4. How to determine protein concentration using a standard curve. 4. How to use the concentration of a dilution to calculate the concentration of the stock solution.
Image of page 1
Electrophoresis and Electroblotting of Proteins 2 10/11/10 3 Extracting Proteins From Cells Why extract proteins from cells? Proteins control virtually all cell activities since they perform all kinds of functions: Enzymes Cytoskeletal proteins Metalloproteins Channels, Pumps G-proteins Motor proteins Receptors Structural Support Molecules Storage proteins 10/11/10 4 Proteins are located in all membranes, the cytoplasm, and the interior of all organelles.
Image of page 2
Electrophoresis and Electroblotting of Proteins 3 10/11/10 In Exercise 6, what was the purpose of DEAE-cellulose in column chromatography? 5 10/11/10 6
Image of page 3
4 7 What kinds of protein will end up in a Crude extract? 8
Image of page 4
Electrophoresis and Electroblotting of Proteins 5 10/11/10 9 Metalloproteins O 2 O 2 O 2 O 2 Hemoglobin Cytochrome C Fe++ 10/11/10 10 Channels and Pumps are Membrane Proteins HYDROPHOBIC HYDROPHOBIC
Image of page 5
Electrophoresis and Electroblotting of Proteins 6 10/11/10 11 How to keep a Protein Structure Intact During Extraction 10/11/10 12 FOUR LEVELS OF PROTEIN STRUCTURE PRIMARY: Ala-Ser-Glut-Val-Tyr-Met-Trp-Phe SECONDARY
Image of page 6
Electrophoresis and Electroblotting of Proteins 7 10/11/10 13 TERTIARY: A B C D 10/11/10 14 QUATERNARY STRUCTURE:
Image of page 7
Electrophoresis and Electroblotting of Proteins 8 10/11/10 15 Extraction Conditions Must Maintain the Protein in its Native Conformation and (optimally) Preserve its Activity.
Image of page 8
Image of page 9

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture