Quantifying 4-HNE in RPE Lysates

RPE lysates were isolated, as described above, at the indicated time points and immediately prepared for 4-HNE analysis. Cleared RPE lysates obtained by centrifugation at 2000× g for 3 min were used in twofold series dilutions. Samples were analyzed using 4-HNE-ELISA kit from Cell Biolabs, San Diego, CA, United States according to the manufacturer’s directions using a Multiskan MCC plate reader (Thermo Fisher Scientific, Waltham, MA, United States) [27 (link)]. Protein was quantified using Bradford reagent (Thermo Fisher, Waltham, MA, USA).

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Dhingra A., Sharp R.C., Kim T., Popov A.V., Ying G.S., Pietrofesa R.A., Park K., Christofidou-Solomidou M, & Boesze-Battaglia K. (2021). Assessment of a Small Molecule Synthetic Lignan in Enhancing Oxidative Balance and Decreasing Lipid Accumulation in Human Retinal Pigment Epithelia. International Journal of Molecular Sciences, 22(11), 5764.

Publication 2021

4-HNE-ELISA kit Multiskan MCC plate reader Bradford reagent

Cell Centrifugation Dilutions Elisa Protein

Corresponding Organization : University of Pennsylvania

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Variable analysis

independent variables

Time points

dependent variables

4-HNE levels

control variables

Centrifugation at 2000× g for 3 min
Bradford reagent for protein quantification

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