The reduced scattering coefficient was specified by linearly scaling the μ ′ s of the IL 20% stock solution, given in [ 24] for the selected IL dilutions.
Moreover, the R S F o data show substantially different spectral responses for measurements of phantoms with different IL dilutions with the same fiber diameter.
Experimental and simulation data are used to identify and investigate the spectral response of reflectance measured in a wide range of IL dilutions as measured by fiber optic device geometries with 1) overlapping source and detector with a range of fiber diameters, and 2) separate source and detector fibers with a wide range of source-detector separations.
50 μg/well underwent electrophoresis on a 12% sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) mini gel (Pierce, Rockford, IL) after dilution in 1× Laemli buffer prior to denaturation at 95°C for 10 minutes.
Zip7 complexes were detected with rabbit α-Zip7 (1:500; ProteinTech, Chicago, IL) in dilution buffer (1% BSA, 0.03% Triton-X100 in PBS) and goat-α-rabbit IgG AlexaFluor 647 (1:500, Life Technologies).
The same supernatants were also subjected to 12% SDS-PAGE, followed by immunoblotting with Abs for human IL-1β (dilution 1 400), caspase-1 (dilution 1 500), and cathepsin B (dilution 1 500) with an ECL Western blotting kit (Amersham, Little Chalfont, UK).
The sections were then incubated with goat anti-human IL-22 (dilution 1 50; R&D Systems) overnight at 4 °C, with a biotinylated mouse anti-goat secondary antibody for 30 min, and with peroxidase-conjugated streptavidin for 10 min.
After microcentrifugation, the supernatant was precleared with blocked protein G plus (Pierce, Rockford, IL, USA) diluted 1 : 10 with dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-Hcl pH 8.1, 167 mM NaCl, 0.5 mM PMFS, 100 ng/ml leupeptin, 100 ng/ml aprotinin), and divided into aliquots.
Sections were incubated overnight at 4°C with mouse monoclonal primary antibodies against human IL-1Ra (1 200 dilution, R&D Systems, Abingdon, UK), IL-1RI (1 50 dilution, R&D Systems), and goat polyclonal primary antibodies against human IL-1α (1 300 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA), IL-1β (1 300 dilution, SantaCruz), and ICE (1 10 dilution, SantaCruz).
