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10.3233/bir-1985-22205 164 VadalaG.RussoF.AmbrosioL.Di MartinoA.PapaliaR.DenaroV

Salivary immune proteins monitoring can help detection of binge and chronic alcohol drinkers: Preliminary findings

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(15) (15) Preparation Fmoc-Gly-Glu(otBu)-Pro-Pro-Pro-Gly-Lys(Boc)-Pro-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin Fmoc-Gly-Glu(otBu)-Pro-Pro-Pro-Gly-Lys(Boc)-Pro-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin (14) In step (14) of Fmoc-Glu(otBu)-Pro-Pro-Pro-Gly-Lys(Boc)-Pro-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin(20DMF)5203DMF3DMF2Fmoc-Gly-OH0.8gDIEA0.4mlTBTU0.56gHOBT0.2gDMF10ml1.53DMF3DMF2Fmoc-Gly-Glu(otBu)-Pro-Pro-Pro-Gly-Lys(Boc)-Pro-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin In Fmoc-Glu(otBu)-Pro-Pro-Pro-Gly-Lys(Boc)-Pro-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin, add decapping reagent (20 %piperidine in DMF solution), reacted at normal temperature for 5 minutes, drained, added capping agent and reacted at normal temperature for 20 minutes, washed 3 times with isopropanol, washed 3 times with DMF, washed 2 times with DMF evaporated, drained, Add 0.8g of Fmoc-Gly-OH, 0.4ml of DIEA, 0.56g of TBTU, 0.2g of HOBT, and 10ml of DMF to react at room temperature for 1.5 hours, dry it with a vacuum pump, wash 3 times with isopropanol, and wash 3 times with DMF

The cell pellet was resuspended and incubated for 30 min on ice with 25 L of staining mix in PBS containing 0.04% BSA, TruStain FcX Block (BioLegend 101320), and the mouse cell surface protein antibody panel containing the following oligo-conjugated anti-mouse antibodies (TotalSeq-A BioLegend) diluted 1:500: CD23 (Cat