Sea water temperature affects mucin O-glycosylation in Atlantic salmon (Salmo salar)

Responsible:

Kristina Thomsson Hulthe . Institute of Biomedicine, University of Gothenburg . kristina.thomsson@medkem.gu.se

Sample preparation

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1.Sample Origin

General information:
Proteins from Atlantic salmon mucus

Growth/harvest conditions for recombinantly produced material:
NA

Treatments and/or storage conditions:
NA

For commercial material, vendor and applicable item information:
NA

Generation of chemically derived material:
NA

2. Sample Processing

Purification steps:
After reductive b elimination, salts are removed by cation exchange beads packed in zip tip pipets, eluted with water, samples were lyphilized and borate complexes removed by repeated MeOH addition/evaporation

2. Defined Sample

Sample name:
O-linked oligosaccharides

HPLC

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1. General settings

Instrument: Agilent 1100 Series

Manufacturer url: https://www.agilent.com/home

Injector: Pal systems - HTC Pal

Injector settings: 3 ul drawn injected into a 2 ul loop

2. Column

Manufacturer: In-house

Model: NA

Type of Chromatography: Graphitized carbon

Type of Material: Graphitized carbon

Column diameter: 250 um

Column length: 10 cm

Particle size: 10 u

Manufacturer url: NA

3. Method run

Temperature: RT

Solvent a: 10 mM (NH4)HCO3

Solvent b: 10 mM (NH4)HCO3 in 80% acetonitrile

Other solvent: NA

Flow rate: approx 3-5 ul/min (static split from 250 ul/min)

Flow gradient: 0-50% B in 40 min, 50-100% B from 40-48 min

Run time: 78 min

Phase: Reversed phase

MS

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N/A