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3 protocols using xylene

1

Optimized Mass Spectrometry Sample Preparation

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Water (HPLC and ULC/MS grade), ethanol, acetonitrile (ACN), trifluoroacetic acid (TFA), and xylene were obtained from Biosolve BV (Valkenswaard, The Netherlands). α -Cyano-4-hydroxycinnamic acid (CHCA), citraconic anhydride, and potassium sulfate were obtained from Sigma Aldrich (St. Louis, MI, USA). Hydrochloric acid fuming 37% was obtained from Supelco (Bellefonte, PA, USA). Basic N-glycan imaging kit was obtained from GlycoPath Inc. (Charleston, SC, USA).
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2

Mass Spectrometry Sample Preparation Protocol

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Water (HPLC and ULC/MS grade), ethanol, acetonitrile
(ACN), trifluoroacetic acid (TFA), and xylene were obtained from Biosolve
B.V. (Valkenswaard, The Netherlands). α-Cyano-4-hydroxycinnamic
acid (CHCA), citraconic anhydride, potassium sulfate, 2,5-dihydroxybenzoic
acid (DHB) 98%, and formic acid (FA) >95% were obtained from Sigma-Aldrich
(St. Louis, MIA). Hydrochloric acid fuming 37% was obtained from Supelco
(Bellefonte, PA). Trypsin was obtained from Promega (Sequencing Grade
Modified Trypsin, Madison, WI). Hematoxylin and Entellan were obtained
from Merck (Darmstadt, Germany). Eosin Y was obtained from J.T. Baker
(Center Valley, PA).
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3

Immunohistochemical Staining Protocol

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Cyanoacrylate (Bison, Goes, the Netherlands) was bought in a local shop. Xylene was purchased from Biosolve (Valkenswaard, the Netherlands), 4% buffered formaldehyde was acquired from Added Pharma (Oss, the Netherlands), paraffin, haematoxylin, and eosin were obtained from Klinipath (Duiven, the Netherlands). DMEM, Ham's F12, and penicillin/streptomycin were purchased from Fisher Scientific (Waltham, MA, USA). Bovine serum albumin, sodium bromide, ethanol, acetone, trypsin, trypsin inhibitor, selenious acid, hydrocortisone, isoproterenol, L-carnitine, L-serine, insulin, α-tocopherol acetate, vitamin C, arachidonic acid, linoleic acid, and palmitic acid were purchased from Sigma-Aldrich (Zwijndrecht, the Netherlands).
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