3 3 5 5 tetramethylbenzidine tmb membrane substrate

A commercially available antibody array kit (Proteome Profiler, Mouse cytokine Array Panel A, R & D Systems) which was coated with 40 capturing antibodies in duplicate on a nitrocellulose membrane (dot blot) was used. The kit contained all the reagents for the assay and was performed as per the manufacturer’s instructions. This cytokine and chemokine antibody array was used to determine the effects of GONPs exposure on cytokine and chemokine by RAW 264.7 macrophage cells.The assay required 500 μL of cell culture supernatants (unstimulated 0 μg/mL GONPs, LPS stimulated 0 μg/mL GONPs and unstimulated 15.6 μg/mL GONPs). Membranes were subjected to an ultra-sensitive chromogenic 3,3′,5,5′-Tetramethylbenzidine (TMB) membrane substrate (Thermo Scientific, Waltham, MA, USA) to reveal sample-antibody complexes labeled with streptavidin-HRP. Photographs were taken of the blots after the exposure to the substrate.

A commercially available antibody array kit (Proteome Profiler, Mouse cytokine Array Panel A, R & D Systems) which was coated with 40 capturing antibodies in duplicate on a nitrocellulose membrane (dot blot) was used. The kit contained all the reagents for the assay and was performed as per the manufacturer’s instructions. This cytokine and chemokine antibody array was used to determine the effects of CD exposure on cytokine and chemokine synthesis by RAW 264.7 macrophage cells. The assay required 500 μL of cell culture supernatants (unstimulated containing 0 μg/mL CDs, LPS stimulated containing 0 μg/mL CDs, and unstimulated containing 500 μg/mL CDs). Membranes were subjected to an ultra-sensitive chromogenic 3,3′,5,5′-Tetramethylbenzidine (TMB) membrane substrate (Thermo Scientific, Waltham, MA, USA) to reveal sample–antibody complexes labeled with streptavidin-HRP. Photographs were taken of the blots after the exposure to the substrate.