Purification of lactate Dehydrogenase

distribution of each enzyme in each fraction (Watson, 2016). The first enzyme marker used was Succinate Dehydrogenase which catalyses the oxidation of succinate to fumarate in the tricarboxylic acid cycle (Hollywood, et al., 2010). This cycle occurs within mitochondria and succinate dehydrogenase is specifically found within the inner mitochondrial membrane. Therefore, succinate dehydrogenase activity is an indicator of mitochondrial activity (Berg, et al., 2007). Mitochondria are large organelles

Isolation and Characterization of Avian Lactate Dehydrogenase To whom correspondence should be addressed Sylvia DaoudKinze and James Proestes, Department of Biochemistry, Portland State University Professor, Portland Oregon, 97207-0751; E-mail: sylviakinzie@gmail.com and proestoj@gmail.com ------------------------------------------------- Keywords: Lactate, Dehydrogenase, Avian, Bradford Assay, Affinity Column Background: Lactate Dehydrogenase also known as LDH is an important NADH dependent

Introduction: Protein purification is a process that can be employed to separate a single protein from a larger starting material which may be anything from an organ to a cell. Isolating a purified protein from a larger fraction enables further analysis such as determination of amino acid sequence, potential biological function, and even evolutionary relationship. (Cuatrecasas 1970) In this experiment, the enzyme lactate dehydrogenase will be purified, this enzyme is found extensively

PURIFICATION OF LACTATE DEHYDROGENASE FROM CHICKEN MUSCLE TISSUE Abstract The enzyme lactate dehydrogenase (LDH) catalyzes the last step of anaerobic glycolysis that is important for the normal function of the body. Purification of LDH is essential to understand its structure and function. The purpose of this experiment was to extract and purify LDH enzyme from chicken muscle tissue using a variety of various. Analytical methods such as activity and protein assay were employed to determine

ligand affinity chromatography was given after the interactions that took place between Blue Dextran, a Cibaron Blue and dextran conjugate, which is used as a void marker in size-exclusion chromatography, and particular kinases. Until then, only purification of various proteins by size-exclusion chromatography with Blue Dextran, like for example, erythrocyte pyruvate kinase, phosphofructokinase, glutathione reductase,

This is seen in the gel electrophoresis where there is a greater number of bands for dye 9-11 than there is for the IEX tubes. The fold purification was expected to be between 3-7-fold as found on the “BRENDA web page” (2017), for this experiment a 1.4-fold (492404/349710) increase in activity was seen after dye ligand chromatography. The retrospective yield was like cation exchange which also

was obtained, precisely minced to exclude extra fat, and then blended with pH 7.2 phosphate buffer. The purpose of blending the tissue with the buffer was to pulverize the cells, causing them to release their contents evenly—most importantly lactate dehydrogenase (LDH)—into the solution. Since other cell components such as proteases which reduce LDH were also released from the lysed cells, the slurry was kept on ice to minimize their kinetic activity. The solution was then centrifuged, creating a pellet