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PWS

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by

Tobias Nijssen

on 7 March 2013

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Transcript of PWS

Results of staining
Choosing
Cutting
Trypsin
Ready peptide chains for LC-MS Usage: Recording retention time and sorting ions
Content: Mobile phase, stationary phase and ...

Small amounts, efficient Mother love thrives on phosphor Analysis of phosphorylated proteins using SDS-PAGE and LC-MS Tobias Nijssen & Tjerk Vredebregt
6V Dalton Den Haag Introduction Main research question: - Milk & precipitation Why do Calcium ions (Ca2+) not form a precipitation in (mother)milk although it is poorly soluble in water? What is phosphorylation and how does it relate to proteins?
How can we analyse proteins to detect phosphorylation? Sub research questions: Milk Electrophoresis/SDS-PAGE LC/MS SDS-PAGE Application: DNA research, Analysis molecules
Working: negative charged particle through gel
- Markers: known substances
- Depending on mass
Staining Coomassie- and silverstaining Making milk Bovine Serum Albumin (BSA) Ovalbumin (OVA) Beta-casseine (B-Cas) 66 kDa 45 kDA 25 kDa Liquid Chromatographer SDS-PAGE Analysis Mass Spectrometer - Charged particles
- Magnetic field, Lorentz force
- Inertia ion
- Detection path of the ion Casting
Layout
Marker
Running Processing and preparing gels Staining Coomassie Silver ions Rough, but easy Very precise, but more complicated Processing LC-MS results 1 Da = 1 u = 1 g/mol Milk Conclusion Serine 49 and phosphoserine 49
Micelle Hypothesis? Our hypothesis: We assume that the phosphorylation of a given amino acid in milk prevents the Ca2+ from precipitating in the milk by capsuling it in one way or another (forming micelles). Accepted Milk contains:
Fats
Sugars (lactose)
Proteins
Calcium Let's begin! And the title? Main research question: Why do Calcium ions (Ca2+) not form a precipitation in (mother)milk although it is poorly soluble in water? The End Are there any questions? We're nearly there!
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