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Technical Note

Assessing AST Released in a Cell Culture Model of Liver Toxicity Using AlphaLISA

Introduction

Aspartate transaminase (AST) or aspartate aminotransferase, also known as AspAT/ASAT/AAT or serum glutamic oxaloacetic transaminase (SGOT), is an important enzyme in amino acid metabolism. AST catalyzes the reversible transfer of an a-amino group between aspartate and glutamate. The enzyme is found in metabolically active organs and tissues, such as the liver, heart, skeletal muscle, kidneys, brain, and red blood cells. Inflamed or injured liver cells leak higher than normal amounts of AST, and other enzymes, resulting in elevated levels found in the bloodstream. A blood test for AST is commonly used as one measure in the detection of liver damage.

In this technical note we demonstrate here the utility and benefits of using AlphaLISA assays for identifying and quantifying levels of AST protein present in cellular lysate and released into the supernatant from a human hepatoma cell line. The data illustrate the benefits of using AlphaLISA assays as a fast, powerful, homogeneous platform for screening modulators of AST expression and release in cultured human cells. The technology has a number of distinct advantages including high signal to background, wide dynamic range, and an extremely simple, straight-forward, no-wash protocol. Download our technical note to learn more.