Glucose Quantification Assay

Introduction

This assay provides a simple and sensitive method to quantify glucose levels from Organ-Chip effluent samples.

Barrier Function Readout Analysis

Introduction

The maintenance or disruption of tissue barriers is an essential part of the pathophysiology of many diseases. The ability to quantitatively characterize tissue barrier is critical in the evaluation of barrier integrity and function.

This protocol is to be used to assess the permeability of an Organ-Chip’s endothelial-epithelial barrier. Apparent permeability (Papp) of tracer molecules is determined by dosing the inlet of one channel, collecting the effluent of both channels, and calculating the amount of compound that crossed through the membrane over time. See full method below and associated Papp Calculator (EC004) for data analysis.

ATP Quantification

Introduction

The CellTiter-Glo® assay kit allows for the quantification of intracellular ATP levels. This endpoint can be used as a marker of cell viability and used in toxicity studies as a measure of cytotoxicity.

Human Cleaved Caspase-3 (Asp 175) Quantification

Introduction

The Cleaved Caspase-3 (Asp175) SimpleStep ELISA® kit is designed for the quantitative measurement of Active Caspase-3 (Asp175) protein in human cells. Caspase-3 is a cytoplasmic cysteine protease involved in the activation cascade of caspases responsible for cell apoptosis.

The SimpleStep ELISA uses an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocaptures the sample analyte in solution. This three-part complex is then immobilized via immunoaffinity of an anti-tag antibody coated on the well.

This endpoint can be applied to measure apoptosis in Organ-Chips as part of toxicity testing or other types of studies.

Gsh-Glo™ Glutathione Assay

Introduction

The GSH-Glo™ Assay is a luminescent-based assay for the detection and quantification of glutathione (GSH), an antioxidant that can prevent damage to cellular components caused by reactive oxygen species such as free radicals, peroxides, and lipid peroxides. GSH is involved in the detoxification of both xenobiotic and endogenous compounds. A change in GSH levels can be used as an indicator of toxicity.

Glycogen Quantification Assay

Introduction

This protocol covers quantification of glycogen on our Organ-Chips.

Triglycerides Quantification Assay

Multiplex Cytokine Assay

Introduction

Goal: Detect and measure secreted cytokines and chemokines from Emulate Organ-Chip effluent samples.

LDH (Cytotoxicity) Assay

Introduction

Goal: Quantify lactate dehydrogenase (LDH) levels from Emulate Organ-Chip effluent samples.

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.