Teacher resources and professional development across the curriculum

Teacher professional development and classroom resources across the curriculum

Monthly Update sign up
Mailing List signup
Search
Follow The Annenberg Learner on LinkedIn Follow The Annenberg Learner on Facebook Follow Annenberg Learner on Twitter
Rediscovering Biology Logo
Home
Online TextbookCase StudiesExpertsArchiveGlossarySearch
Online Textbook
Back to Unit Page
Unit Chapters
Genomics
Proteins & Proteomics
What is Proteomics?
Introduction to Protein Structure
Determining Protein Structure
Structure and Function Relationships of Proteins
Protein Modification
Genomics-Based Predictions of Cellular Proteins
2D Gel Electrophoresis to Identify Cellular Proteins
Mass Spectrometry to Identify Cellular Proteins
Identifying Protein Interactions
The Yeast Two-Hybrid System
Protein Microarrays
Protein Networks
Proteomes in Different Organisms
Proteomics and Drug Discovery
Ethics and the Economics of Drug Discovery
Evolution & Phylogenetics
Microbial Diversity
Emerging Infectious Diseases
HIV & AIDS
Genetics of Development
Cell Biology & Cancer
Human Evolution
Neurobiology
Biology of Sex & Gender
Biodiversity
Genetically Modified Organisms
2D Gel Electrophoresis to Identify Cellular Proteins

While computer-based methods are powerful, they can only predict the function of proteins for which some information is already available. How do we understand the proteins that we don't already know about? This requires experimental approaches.

One way to identify proteins is to extract all the proteins from a sample of cells and separate them in a gel matrix, using a technique called polyacrylamide gel electrophoresis (PAGE). The proteins are separated by size, with the smaller proteins moving faster through the gel than the larger proteins. After staining, a pattern of bands appears that corresponds to the proteins in the cell. However, this technique can only resolve a few hundred proteins, and cannot separate proteins of very similar size.

Figure 4. 2D gel of proteins
A modification of this procedure - called 2D gel electrophoresis - separates proteins into two dimensions, using two different characteristics. Proteins are separated in the first dimension by their isoelectric point (pI), the specific point at which the net charge of the protein is zero. These separated proteins, in a flat gel strip, are then placed on a standard polyacrylamide gel. Every protein band that was separated in the first dimension according to its isoelectric point is now separated in the second dimension by its size. The result is small spots, each representing a protein; even proteins of the same size will be resolved if they have a different isoelectric point. A good 2D gel can resolve one thousand to two thousand proteins, which appear, after staining, as dots in the gel (Fig. 4). This technique is useful when comparing two similar samples to find specific protein differences; for example, comparing the proteins in a tumor cell versus a normal cell. However, it can miss very small proteins or non-abundant proteins.

Back Next


  Home  |  Catalog  |  About Us  |  Search  |  Contact Us

| Follow The Annenberg Learner on Facebook
 

  © Annenberg Foundation 2013. All rights reserved.
Privacy Policy