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2 protocols using anti bsa rabbit igg

1

Quantifying Immune Cell Responses to LPS Stimulation

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Lipopolysaccharide (LPS) from Escherichia coli O111:B4 (Sigma-Aldrich, St. Louis, MO, USA) was used. Phorbol 12-myristate 13-acetate (PMA), ionomycin, bovine serum albumin (BSA), anti-BSA rabbit IgG and anti-RASAL3 rabbit IgG were also purchased from Sigma-Aldrich. SYTOXRed, DAPI (4’,6-diamidino-2-phenylindole) and CellROX™Green were purchased from Thermo Fisher Scientific (Waltham, MA, USA). A Cytofix/Cytoperm kit with GolgiStop™ was purchased from BD Bioscience (Franklin Lakes, NJ, USA). Anti-CD45 (30-F11), anti-CD11b (M1/70), anti-Ly6G (1A8), anti-CD16/CD32 (2.4G2) (93) and PE donkey anti-rabbit IgG (Poly4064) were all purchased from Biolegend (San Diego, CA, USA). The isotype-matched control for each antibody was purchased from the same manufacturers.
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2

Nanoimprinted Optofluidic Biosensor Protocol

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Sodium phosphate buffer (PBS, 8 mM Na 2 HPO 4 , 2 mM, 137 mM NaCl, 2.7 mM KCl, pH 7.4), PBS-T (PBS with polysorbate 20 0.05% v/v), were prepared with purified water (Milli-Q, Millipore Iberica, Darmstadt, Germany) and filtered through 0.2 μm polyethersulfone membranes (Merck, Darmstadt, Germany). Polydimethylsiloxane (PDMS) Sylgard 184 was from Dow Corning (Wiesbaden, Germany). Bovine serum albumin (BSA), polysorbate 20 (Tween 20), antiBSA rabbit IgG, C-reactive protein (CRP), casein and human serum (human male, AB plasma) were supplied by Sigma-Aldrich (Madrid, Spain). Single-mode optical fibers SMF-28 were purchased from Corning (Madrid, Spain). The silicon grooved nanostructure (555.5 nm period, 140 nm groove depth, duty cycle 50%) used as a master to prepare the micro-contact printing stamp, was from LightSmyth (Eugene, OR, USA).
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