4. Effluent Analysis

Total ROS and RNS Assay

Introduction

Accumulation of free radicals, coupled with an increase in oxidative stress, has been implicated in the pathogenesis of several disease states and in the mechanism of action for toxicity of many compounds. This protocol can be used to measure these toxicity endpoints. The OxiSelect™ In Vitro ROS / RNS Assay Kit allows for the measurement of the total amount of free radicals in cell effluent by using a specific ROS / RNS fluorogenic probe. The fluorescence intensity is proportional to the total ROS / RNS levels within the sample.

This protocol uses the Proximal Tubule Kidney-Chip as a reference point. These methods and assay conditions could change with different Organ-Chips.


⃪ Back to Protocols

Creatinine Quantification Assay

Introduction

Creatinine, a non-protein nitrogenous (NPN) waste product, is produced from the breakdown of creatine and phosphocreatine. Creatinine levels can be used as an indicator of renal function.

To measure creatinine secretion in the Proximal Tubule Kidney-Chip, the vascular channel (bottom channel) is perfused with medium containing 1mg / dL of human creatinine, while the secretion of creatinine in the epithelial channel (top channel) is measured from chip effluent that collects in outlet reservoirs of the Pod™ portable module.

This protocol uses the Proximal Tubule Kidney-Chip as a reference point. These methods and assay conditions could change with different Organ-Chips.


⃪ Back to Protocols

Keratin 18 (K18) Assay

Introduction

Keratin 18 (K18) is an intracellular protein expressed at high levels by many types of epithelial cells. During cell death, the cellular content of soluble K18 will be released into the extracellular compartment. The M65 EpiDeath® ELISA measures soluble keratin 18 (K18) (cytokeratin 18 [CK18]) released from dying cells and can be used in the research of overall cell death (due to apoptosis and necrosis) of epithelial cells.


⃪ Back to Protocols