Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags. |
-Run a standard curve with each assay.
-Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
-Do not cover plate with foil. |
-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity.
-Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days. |
Upstream tips |
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags. |
Protocol tips |
-Run a standard curve with each assay.
-Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
-Do not cover plate with foil. |
Downstream tips |
-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity.
-Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days. |
Publication protocol
Bone marrow cells were flushed and dispersed to single cell suspension with phosphate-buffered saline (PBS; Ca2+/Mg2+ free). After red blood cell lysis in 20 mL 0.2% NaCl for 20 seconds and neutralized in 20 mL 1.8% NaCl, cells were pelleted down and subjected to gradient centrifugation in 6 mL 1119/1077 Histopaque (Sigma). Neutrophils were collected from the interface and used for in vitro studies. Neutrophils were stimulated with either lipopolysaccharides (LPS) (50 ng/mL) or Escherichia coli (10 multiplicity of infection [MOI]) for 30 minutes. Matrix metallopeptidase 9 (MMP-9) and myeloperoxidase (MPO) in serum and condition medium were measured by mouse MPO enzyme-linked immunosorbent assay (ELISA) Kit (Thermo Fisher, EMMPO) and mouse pro-MMP-9 ELISA Kit (Thermo Fisher, EMMMP9).
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