Use of Enzymatic Biosensors to Quantify Endogenous ATP or H<sub>2</sub>O<sub>2</sub> in the Kidney

Oleg Palygin; Vladislav Levchenko; Louise C. Evans; Gregory Blass; Allen W. Cowley Jr.; Alexander Staruschenko

doi:10.3791/53059

Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

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Use of Enzymatic Biosensors to Quantify Endogenous ATP or H₂O₂ in the Kidney

Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

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10:00 min

October 12, 2015

DOI: 10.3791/53059-v

Oleg Palygin¹, Vladislav Levchenko¹, Louise C. Evans¹, Gregory Blass¹, Allen W. Cowley Jr.¹, Alexander Staruschenko¹

¹Department of Physiology,Medical College of Wisconsin

Enzymatic microelectrode biosensors enable real-time measurements of extracellular cell signaling in biologically-relevant concentrations. The following protocols extend the applications of biosensors to the ex vivo and in vivo detection of ATP and H₂O₂ in the kidney.

The overall goal of these procedures is to measure real-time endogenous interstitial concentrations of a TP and hydrogen peroxide in whole kidneys. This is accomplished by first rehydrating and calibrating sensors for ex vivo and in vivo applications to known concentrations of a TP and hydrogen peroxide. For ex vivo sensor installation, the kidney was surgically instrumented and cleaned from the blood.

This procedure is performed by flushing the kidney through the aorta. Then the rat kidney is surgically isolated, placed in a recording chamber and perfused with saline. A TP and hydrogen peroxide are measured using ex vivo sensors for the in vivo procedure.

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