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2 protocols using rituximab rtx

1

Multicolor Flow Cytometry Antibody Labeling

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Antibodies were diluted 1:100 on 1 × 106 cells for flow cytometry; Cytotect (Biotest); αCD107a-APC (BD FastImmune clone H4A3); αCD56-BV650 (Biolegend clone 5.1H11); αCD3-FITC (Biolegend clone UCHT1); αhuman-IgG-PE (BD); human Fcγ-TexasRed (Rockland); rituximab (Rtx, Roche); herceptin (Hc, Roche); αhuman-IgG-PE (Miltenyi Biotec); αHis-PE (Miltenyi Biotec); αCD20-PE (Miltenyi Biotec); polyclonal rabbit-αhuman IgG-FITC (ThermoFisher); THE Anti-His-HRP (Genscript); MSL-109 anti HCMV gH (Absolute Antibody).
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2

Papain Activity Assay Protocol

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Butyl acrylate, glycidyl methacrylate, trimethylolpropane triacrylate (TMPTA), potassium persulfate, N,N,N′,N′-tetramethylethylenediamine (TEMED), methanol, glycine, Nα-benzoyl-L-arginine ethyl ester (BAEE), Nα-benzoyl-DL-arginine 4-nitroanilide hydrochloride (BAPNA), papain from papaya latex, Trizma® base (Tris), and ethylenediaminetetraacetic acid (EDTA) were purchased from Sigma-Aldrich (Milan, Italy). Synperonic PE/L121 was kindly provided by Croda Italiana SpA (Mortara, Italy). Tetrahydrofuran (THF), acetonitrile, and L-cysteine were from PanReac AppliChem ITW Reagents (Cinisello Balsamo, Italy). Potassium dihydrogen phosphate and formic acid were from Carlo Erba (Cornaredo, Italy).
Deionized water was obtained from a Milli-Q® Integral purification system from Merck KGaA (Darmstadt, Germany).
Rituximab (RTX) was purchased as MabThera from Roche (Basel, Switzerland).
Chromolith® Flash WP300 Epoxy silica columns of 4.6 × 25 mm were kindly provided by Merck KGaA (Darmstadt, Germany), while Omnifit® EZ-Solvent Plus glass columns of 10 × 100 mm by Diba Industries, Ltd., were purchased from Sepachrom-Mega Srl (Rho, Italy).
All reagents were of analytical grade.
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