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2 protocols using anti snap tag antibody polyclonal

1

Labeling Amyloid-beta with Fluorescent Probes

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The pSNAP-tag(m) plasmid, which was used as backbone, is available at Addgene (#101135). The transfection reagent TransIT-LT-1 was purchased from Mirus (Lot81023060) and the ncAA TCO*K (axial trans-cyclooct-2-ene–L–lysine; CAS 1801936-26-4) was purchased from SiChem. We used four fluorescent dyes for labeling. The SNAP-Cell® 647-SiR (S9102 S) and the SNAP-Cell® TMR-Star (S9105 S) were purchased from New England BioLabs and the 6-methyl-tetrazine-BDP-FL is produced by Jena Bioscience (RK011-001). The SiR-DNA kit (SC007) was purchased by Spirochrome. For detecting Aβ, we used the High sensitive IBL-ELISA: Amyloid-beta (1–42) (LOT. 1D-818) and Amplex® UltraRed from Molecular Probes, Thermo Fisher Scientific. The 4–20% polyacrylamide Bis-Tris gels were purchased by BioRad. The cOmplete™, EDTA-free Protease Inhibitor Cocktail was purchased from Sigma (000000005056489001). The primary antibodies we used are the Anti-SNAP-tag® antibody (Polyclonal), (1 : 200) from New England BioLabs (P9310 S) and the Anti-β-Amyloid, 1–16, Clone: 6E10 (1 : 1000) from BioLegend (803001). The secondary antibodies are the IRDye 680RD Donkey anti-Rabbit (926-68073) and IRDye 800 CW Donkey anti-mouse (92632212) from LI-COR Biosciences.
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2

Immunofluorescence Labeling of Cell Membrane Proteins

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We obtained anti-human CCR5 45531 from R and D Systems (Minneapolis, MN), anti-actin from Sigma Aldrich (St. Louis, MO), anti-Giantin 924302 from Biolegend (San Diego, CA), Alexa Fluor 488 FluoroNanogold goat anti-mouse IgG Fab from Nanoprobes.com (Yaphank, NY), AffiniPure Donkey Anti-Rabbit IgG (H + L) from Jackson ImmunoResearch (West Grove, PA), Alexa Fluor 488 goat Anti-Mouse IgG (H + L) from Invitrogen (Carlsbad, CA), Anti-SNAP-tag Antibody (Polyclonal) from New England Biolabs (Ipswichm, MA). Hybridoma cells expressing anti-Myc 9E10 were obtained from ATCC. Saponin (from quillaja bark), LPS (from Escherichia coli 0111:B4), glutaraldehyde, and fumonisin B1 (from Fusarium moniliforme) was purchased from Sigma Aldrich (St. Louis, MO). Recombinant human CCL5 was obtained from R and D Systems (Minneapolis, MN).
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