Bradford assay

Manufactured by GeneCopoeia

Sourced in United States

The Bradford assay is a colorimetric analytical procedure used to measure the concentration of protein in a solution. It involves the binding of Coomassie Brilliant Blue G-250 dye to proteins, which results in a color change that can be measured spectrophotometrically.

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Lab products found in correlation

Protein extraction buffer, by Thermo Fisher Scientific (2 mentions) Quantikine elisa kit, by R&D Systems (2 mentions)

2 protocols using bradford assay

Quantifying Corneal Nox2 and Angiogenic Factors

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The mouse corneas were isolated and weighed. Corneal tissues (four corneas/group from two batches) were harvested and homogenized with protein extraction buffer (Thermo Fisher Scientific). The mixture from each sample was centrifuged at 10,000 g for 3 minutes at 4 °C and the supernatant was collected. Total protein was quantified using Bradford assay (p010, GeneCopoeia, Rockville, MD, USA). Mouse nicotinamide adenine dinucleotide phosphate oxidase 2 (Nox2) ELISA Kit (My BioSource, cat no. MBS269961) was used to quantify Nox2 content in cornea. The angiogenic cytokines (MMP2, MMP9, and VEGF) content in each group was measured using the Quantikine^® ELISA kit and Mouse VEGF/total MMP9/MMP2 Immunoassay (R&D Systems, Minneapolis, MN, USA) in triplicate. The experiments were conducted according to the manufacturer’s protocol.

Chu Y.C., Fang H.W., Wu Y.Y., Tang Y.J., Hsieh E.H., She Y., Chang C.Y., Lin I.C., Chen Y.J., Liu G.S, & Tseng C.L. (2023). Functional Peptide-Loaded Gelatin Nanoparticles as Eyedrops for Cornea Neovascularization Treatment. International Journal of Nanomedicine, 18, 1413-1431.

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Quantifying Corneal VEGF and MMP-9

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Corneal tissues (6 eyeballs/group from two batches) were harvested and homogenized with protein extraction buffer (Thermo Fisher Scientific). The mixture from each sample was then centrifuged, and the supernatant was collected. Total protein of cornea lysate was quantified by Bradford assay (p010, GeneCopoeia, Rockville, MD, USA). An equal amount of total protein (15 µg in 100 µL) from each sample was used to quantify the VEGF and MMP-9 by ELISA (Quantikine ELISA kits, R&D system, Minneapolis, MN, USA). This experiment was conducted according to the manufacturer’s protocol.

Miyagawa T., Chen Z.Y., Chang C.Y., Chen K.H., Wang Y.K., Liu G.S, & Tseng C.L. (2020). Topical Application of Hyaluronic Acid-RGD Peptide-Coated Gelatin/Epigallocatechin-3 Gallate (EGCG) Nanoparticles Inhibits Corneal Neovascularization via Inhibition of VEGF Production. Pharmaceutics, 12(5), 404.

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