Download
Skip this Video
1 / 12
Biochemistry Lab - PowerPoint PPT Presentation
- 60 Views
- Uploaded on
Biochemistry Lab. Protein Purity Assessment and Identification. Coomassie Staining. How does Coomassie interact with proteins?. Coomassie Staining. Benefits. Limitations. Western Blotting. Transfer – Block Apply Primary Antibody Apply Secondary Antibody. Western Blots.
Download Presentation 
An Image/Link below is provided (as is) to download presentation
PowerPoint Slideshow about 'Biochemistry Lab' - garret
An Image/Link below is provided (as is) to download presentation
Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.
While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.
- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
Biochemistry Lab
Protein Purity Assessment and Identification
Coomassie Staining
How does Coomassie interact with proteins?
Soft Ionization Techniques
Matrix Assisted Laser Desorption Ionization (MALDI)
Electrospray Ionization (ESI)
- Aqueous sample introduced to metal capillary
- High voltage (2000-4000 V) applied
- Released to vacuum
- Desolvation of aerosol leaving highly charged ions
- Aqueous sample is cocrystallized on a metal surface with a Matrix
- Intense Laser beam is directed toward sample/matrix mixture - desorption
- Matrix absorbs the energy and is ionized
- Some of the charge is transferred to the analyte
Separation Techniques
Quadrupole
Flight Tube
- Four rods are arranged opposite each other and connected electronically
- Voltage applied to each rod is carefully regulated
- The trajectory of a charged particle is influenced by the electric field
- Molecules separate by the time it takes for them to travel from the ion source to the detector
- Resolution is dependent on tube length (limits resolving power)
- Reflectron enhances the resolution
In lab this week
- Run and stain (coomassie) the gels you prepared last week
- Use Fractions 1-9 and a MW ladder
- MALDI-ToF MS of your fractions
SinapicAcid (SA)
