Cell-based Assays

Acetokinase (ACK) is primarily found in microorganisms and functions as a key enzyme in bacterial carbon metabolism and energy metabolism, playing a central role particularly in the methanogenesis pathway of archaea. ACK catalyzes the conversion of acetate and ATP into acetyl phosphate and ADP. Subsequently, pyruvate kinase catalyzes the formation of ATP and pyruvate from ADP and PEP. Lactate dehydrogenase then catalyzes the reaction that converts pyruvate and NADH into lactate and NAD+. By measuring the rate of NADH oxidation to NAD+ at a wavelength of 340 nm, one can determine the activity of ACK.

Acetyl CoA Carboxylase (ACC) catalyzes the carboxylation of acetyl-coa to malonyl-coa in vivo, and is A key enzyme in the synthesis of fatty acids and many secondary metabolites. ACC activity determines, in part, the rate of fatty acid synthesis and the level of oil content. CheKine™ Micro Acetyl CoA Carboxylase (ACC) Activity Assay Kit can detect animal and plant tissues, bacteria and biological samples such as cells, serum or plasma. In this kit, ACC was able to catalyze acetyl-coa, NaHCO3 and ATP to produce malonyl-CoA, ATP and inorganic phosphorus, and ACC activity was determined by measuring the increase in inorganic phosphorus by ammonium molybdate method.

Polysaccharide are ubiquitous substances in organisms, which are a kind of natural polymers connected by Aldose or ketose through glycosidic bonds. it is an important biological macromolecule in the organism and one of the basic substances to maintain the normal operation of life activities. CheKineTM Micro Total Polysaccharide Content Assay Kit can be used to detect biological samples such as animal and plant tissues. In the kit, the total polysaccharide were extracted by water extraction and alcohol precipitation, and the content of total polysaccharide was determined by phenol-sulfuric acid method.

Proline dehydrogenase (ProDH) is a key enzyme that catalyzes the degradation of proline in mitochondria. Reducing the activity of ProDH is of great significance for regulating osmotic balance, preventing plant damage caused by osmotic stress, scavenging free radicals and protecting cell structure. CheKineTM Micro Proline Dehydrogenase (ProDH) Activity Assay Kit can be used to detect biological samples such as animal and plant tissues,cells or bacteria other liquid samples. In this kit, ProDH catalyzed the dehydrogenation of proline to pyruvic acid, and the removed hydrogen was transferred to reduce 2-dichlorophenol indophenol (DCPIP) by phenazine dimethyl ester sulfuric acid (PMS), and there was a characteristic absorption peak at 600 nm. Through the decrease of 600 nm absorbance, the reduction rate of 2-dichlorophenol indophenol (ProDH) was measured, which represented the activity of DCPIP.

Urea is the end product of nitrogenous compound degradation in living organisms, undergoing conversion to ammonia under the catalysis of urease. Blood urea nitrogen (BUN) is one of the principal indicators of renal function. In a sample, urea nitrogen reacts in a solution containing ferric chloride and phosphate, along with diacetyl monoxime and thiourea, upon heating to produce a red diazine compound. The intensity of the color of this compound is directly proportional to the concentration of urea nitrogen present. The determination of urea nitrogen levels is accomplished using the diacetyl monoxime method, which relies on colorimetric analysis of the reaction mixture to quantify urea nitrogen content in serum or urine specimens.

Polyamine oxidase (PAO) is a key enzyme that catalyzes the oxidation of polyamines in organism. In addition, PAO can regulate the level of polyamines and the concentration of products in organism, and participate in the response to stress and the growth and development process of each plant. CheKine™ Micro Polyamine Oxidase (PAO) Activity Assay Kit can detect biological samples such as Animal and Plant Tissues, Serum or Plasma. In this kit, PAO catalyzed the oxidation of polyamines to produce hydrogen peroxide, which reacts with the substrate in the presence of peroxidase, and the product has a characteristic absorption peak at 500 nm. The activity of PAO is reflected by measuring the rate of increase in absorbance value.

Lipoprotein lipase (LPL) is an enzyme synthesized by adipocytes, cardiomyocytes, skeletal muscle cells, mammary cells, macrophages and other parenchymal cells. LPL catalyzes the hydrolysis of triglyceride to fatty acids and monophosphate for tissue oxidation for energy supply and storage. Hydrolysis of 4-nitrobenzene palmitate by LPL yields 4-nitrophenol with a characteristic absorption peak at 400 nm.

Mammalian CarE also known as aliphatic esterase (aliesterase) is widely distributed in tissue and organism, belonging to the serine hydrolase family. CarE catalyze hydrolysis of endogenous and exogenous substances containing ester bonds, amide bonds, and thioester bonds, but can’t catalyze hydrolysis of acetylcholine and its analogues. CarE take part in lipid transport and metabolism, and related with detoxification and metabolism of many drugs, environmental poisons and carcinogens. Organophosphorus pesticides can bind to CarE and inhibit CarE activity. CarE can catalyze acetic acid-1-naphthalene ester to produce naphthalene ester, solid blue color development. Determination of 450 nm light absorption increase rate could calculate CarE activity.

Pepsin is secreted by the main cells of the gastric mucosa and breaks down proteins in food into small peptides. Generally used for the identification of neuropathic hypoacidosis, chronic gastritis, chronic gastric dilatation, chronic duodenitis and other symptoms will also cause the decrease of pepsin secretion. CheKineTM Micro Pepsase Activity Assay Kit can be used to detect biological samples such as animal tissues. In the kit, pepsase can catalyze the hydrolysis of hemoglobin, and the hydrolysate appears blue after reacting with Folin reagent, and the color of pepsin is proportional to the activity of pepsin in a certain range.