Stress impairs skin barrier function and induces α2-3 linked N-acetylneuraminic acid and core 1 O-glycans on skin mucins in Atlantic salmon, Salmo salar

Responsible:

John Benktander . Gothenburg university . john.benktander@gu.se

Sample preparation

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1.Sample Origin

General information:
O-glycans from Atlantic salmon skin mucus.

Growth/harvest conditions for recombinantly produced material:
NA

Treatments and/or storage conditions:
-80 C

For commercial material, vendor and applicable item information:
NA

Generation of chemically derived material:
NA

2. Sample Processing

Purification steps:
Crude mucin exreaction.

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: HTC PAL

Injector settings: 3 ul

2. Column

Manufacturer: In-house

Model: Hypercarb

Type of Chromatography: NA

Type of Material: Porous graphitized carbon

Column diameter: 0.25

Column length: 100

Particle size: 5

Manufacturer url: NA

3. Method run

Temperature: RT

Solvent a: 10 mM (NH4)HCO3 in water

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

Other solvent: NA

Flow rate: 10 ul/min

Flow gradient: 0-40%

Run time: 40 min

Phase: reverse phase

MS

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1. General features

Date stamp: Wed Dec 30 UTC 2020

Instrument: Thermo Fisher LTQ Linear Ion Trap ESI

Customizations from the manufacturer's specification:
NA

ion_mode: -

2. Ion sources

(a) Electrospray ionisation - ESI

Sprayer type : Fed

Interface name: NA

Other data for the Interface: NA

Sprayer name: IonMax standard ESI source

Other data for the sprayer: Equipped with a stainless steel needle

Relevant voltages: Electrospray voltage of 3.5 kV and capillary voltage of −33.0 V.

Degree of prompt fragmentation evaluated: Yes

In-source dissociation performed: Yes

Other parameters if discriminant for the experiment: NA

(b) MALDI

Plate composition (or type): NA

Matrix composition: NA

Deposition technique: NA

Relevant voltages: NA

Degree of prompt fragmentation evaluated: NA

PSD (or LID/ISD) summary: NA

Operation with or without delayed extraction: NA

Laser type: NA

Other laser related parameters: NA

url: NA

3. Ion transfer optics

Hardware options: NA

3. Post-source componentry

Collision-Induced Dissociation - CID

CID. Gas composition: He

CID. Gas pressure: NA

CID. Collision energy: Collision energy 35%

Electron Transfer Dissociation - ETD

ETD. Reagent gas: NA

ETD. Pressure: NA

ETD. Reaction time: NA

ETD. Reagent atoms: NA

Electron Capture Dissociation - ECD

ECD. Emitter type: NA

ECD. Voltage: NA

ECD. Current: NA

TOF drift tube

TOF. Reflectron status: NA

Ion Trap

Ion trap. Final MS stage achieved: NA

Ion mobility

Ion mobility. Gas type : NA

Ion mobility. Pressure: NA

Ion mobility. Instrument parameters: NA

FT-ICR

Peak selection: NA

FT-ICR. Pulse: NA

FT-ICR. Width: NA

FT-ICR. IR: Decay time: NA

FT-ICR. IR: NA

FT-ICR. Other parameters: NA

Detectors

Detector type: NA