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2 protocols using goat anti human gpnmb

1

Antibody Panel for Neurodegenerative Markers

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The following antibodies were used in this study: mouse anti-Galectin-3 (BioLegend, 126702), goat anti-human GPNMB (R&D Systems, AF2550), goat anti-mouse GPNMB (R&D Systems, AF2330), rat anti-mouse LAMP1 (BD Biosciences, 553792), rabbit anti-IBA-1 (Wako, 01919741), goat anti-AIF-1/Iba1 (Novus Biologicals, NB100–1028), rat anti-CD68 (Bio-Rad, MCA1957), and sheep anti-PGRN (R&D Systems, AF2557), mouse anti-MBP (Millipore, SMI-99), Goat anti-Olig2 (R&D Systems, AF2418), mouse anti-APC (Millipore, OP80), rabbit anti-Perilipin2 (Proteintech Group,15294-1-AP), sheep anti-TREM2 (R&D Systems, AF1729). Detailed information is provided in Supplementary Table 1.
The following reagents were also used in the study: Dulbecco’s modified Eagle’s medium (DMEM)(Cellgro, 10–017-CV), Hanks’ Balanced Salt Solution (HBSS) (Cellgro, 21–020-CV), Dulbecco’s modified Eagle’s medium/Ham’s F-12 (DMEM/F-12)(Cellgro, 10–092-CV), 0.25% Trypsin (Corning, 25–053-CI), Autofluorescence Quencher (Biotium, 23007), Odyssey blocking buffer (LI-COR Biosciences, 927–40000), and O.C.T compound (Electron Microscopy Sciences, 62550–01).
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2

Quantitative Analysis of gpNMB Expression

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gpNMB was stained using goat anti-human gpNMB (R&D Systems) primary antibody (1:500) at 4°C overnight, then with an anti-goat-Dylight 488 (Novus) secondary antibody (1:2500) at ambient temperature for 1 hr. Slides were counterstained with DAPI mounting solution (Sigma). The high-resolution images of tissue slices were acquired using a digital whole slide scanner (Nanozoomer 2-HT, Hamamatsu. Bridgewater, NJ) using a 20×/0.0.75 lens (Olympus, Center Valley, PA). The images were then quantified using the image analysis module of a digital pathology software Visiomorph (VisioPharm, Broomfield, CO).
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