Search for notes by fellow students, in your own course and all over the country.
Browse our notes for titles which look like what you need, you can preview any of the notes via a sample of the contents. After you're happy these are the notes you're after simply pop them into your shopping cart.
Title: Intro Proteomics
Description: Introduction to Proteomics from University.
Description: Introduction to Proteomics from University.
Document Preview
Extracts from the notes are below, to see the PDF you'll receive please use the links above
Proteomics
What is proteomics?
• Study of the protein content of a cell
• What proteins are produced by a specific
tissue
• What new proteins are produced after an
event
• What proteins interact with each other
Methods
• Protein separation techniques
– SDS-PAGE
• Sodium dodecylsulphate polyacrylamide gel
electrophoresis
• Gives approximate size information for proteins
– 2D-gel electrophoresis
• Gives approx size information and pI
– What is the pI?
– Comparative 2D gels
SDS-PAGE
• Denaturing
• Separates by mass
• Can resolve many
proteins
• Can excise and
sequence
• Determine
molecular weight by
Rf calculation
Loading a protein gel
Loading a protein gel
Running a protein gel
Staining the proteins
Example of a real protein gel:
Protein purified by affinity chromatography by
students from 2004 years BC4904 group
• Very powerful 2D-gels
technique
• Separates many
hundreds of
proteins
• Can excise and
sequence
• Gives info on pI
and mass of
proteins
• Used for
comparative
studies
Comparative 2D gels
• Examine cells after an event
Control condition
Test condition
• New spots, and missing spots
How they really look
• Very complex, need computer software to
keep track of all the info
What can you do with this info?
• Search genome sequence if it is available for the
organism
– Have rough info on size and pI
• Excise spot
– Protein sequence
• Older method – Edman degradation
• MS-MS (tandem mass spec)
– Mass determination by Mass Spec (MS)
• Very accurate technique
Mass Spectrometry
Applications:
•
•
•
•
Determine molecular weights
Investigate intermolecular interactions
Sequence proteins and peptides
Identification of proteins
Features:
• Very accurate measure of molecular mass
...
01% accuracy
...
Two methods used to produce the ions
MALDI and ES
Matrix-Assisted Laser Desorption-Ionization
(MALDI)
Laser
Analyte
TOF analysis
Matrix
Embedded sample
• Biological samples have low
vapour pressure
• Implant in matrix which
absorbes UV
• Pulse with energy from laser
• Generates ions
MALDI-TOF analysis
New methods using MS
• The MS/MS analysis can
– Detect novel peptides in one sample
– Relative abundance of peptides
– Sequence information
How does a protein behave
• Many perform
reactions, pass
signals, bind to other
proteins
• Function genomics
looks at this
– Structural analysis
– Biochemical analysis
– Networks of
interactions
Yeast protein interaction network
Title: Intro Proteomics
Description: Introduction to Proteomics from University.
Description: Introduction to Proteomics from University.