Anti-ISG15 rabbit serum, gift of Klaus-Peter Knobeloch, was used as primary antibody for immunoblotting. Secondary antibodies for immunoblotting were from KCL: goat anti-rabbit HRP (474-1506) and goat anti-mouse HRP (474-1806), used for the IgG heavy chain blot. Fluorescently labelled antibodies for cytofluorometry (FACS) against Ly6C (clone HK1.4), CD11b (clone M1/70), I-A/I-E (clone M5/114.15.2), CD11c (clone N418), Ly6G (clone 1A8), F4/80 (clone BM8), CD103 (clone 2E7), CD8α (clone 53-6.7), CD45R/B220 (clone RA3-6B2), CD49b (clone DX5), TCRβ (clone H57-597) and purified CD16/32 (anti Fc-γ receptor III/II, clone 93) were purchased from Biolegend. Anti-mouse IL-1β (clone NJTEN3), anti-mouse IL-12 (clone C15.6) and Rat IgG1 k isotype control (clone eBRG1) were purchased from eBioscience. Anti-FITC MACS beads (130-048-701) used for negative selection for the intracellular staining were from Miltenyi Biotec. ELISA Pierce TMB substrate kit (34021) was from Thermo Fisher Scientific. Dulbecco’s PBS was from VWR (21-031-CV).
Dulbecco s pbs
Manufactured by Avantor
Dulbecco's Phosphate Buffered Saline (PBS) is a balanced salt solution commonly used in biological research. It maintains the osmotic balance and pH of cell cultures and other biological samples. Dulbecco's PBS provides a physiologically relevant environment for various cell-based assays and experiments.
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2 protocols using dulbecco s pbs
1
Immunoblotting and FACS Antibody Validation
2
Immunoblotting and Flow Cytometry Analyses
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Anti-ISG15 rabbit serum, a gift from K.-P. Knobeloch, was used as primary Ab for immunoblotting. Secondary Abs for immunoblotting were from KPL: goat anti-rabbit HRP (474-1506) and goat anti-mouse HRP (474-1806), used for the IgG H chain blot. Fluorescently labeled Abs for FACS against Ly6C (clone HK1.4), CD11b (clone M1/70), I-A/I-E (clone M5/114.15.2), CD11c (clone N418), Ly6G (clone 1A8), F4/80 (clone BM8), CD103 (clone 2E7), CD8α (clone 53-6.7), CD45R/B220 (clone RA3-6B2), CD49b (clone DX5), TCRβ (clone H57-597), and purified CD16/32 (anti Fc-γ receptor III/II, clone 93) were purchased from BioLegend. Anti-mouse IL-1β (clone NJTEN3), anti-mouse IL-12 (clone C15.6), and Rat IgG1 k isotype control (clone eBRG1) were purchased from eBioscience. Anti-FITC MACS beads (130-048-701) used for negative selection for the intracellular staining were from Miltenyi Biotec. ELISA Pierce TMB substrate kit (34021) was from Thermo Fisher Scientific. Dulbecco’s PBS was from VWR (21-031-CV).
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