Cell-based Assays

The RayBiotech thiol assay kit is designed based on the Ellman’s reaction, using DTNB reagent, and optimized for the high-throughput microtiter form. Quickly measure the amount of total free thiol in a given sample with a range of 0-40 nmole per well.

Glucose-6-phosphate (6PG), also known as Glucose 6-phosphate, is an intermediate product of glycolysis and pentose phosphate pathway, which is widely found in animals, plants and microorganisms. In the first step of glycolysis, glucose is catalyzed by hexokinase to form glucose-6-phosphate, which is then catalyzed by phosphate glucose isomerase to form fructose-6-phosphate to continue the other steps of glycolysis. However, in the pentose phosphate pathway, 6PG is its first substrate, and this process is also the main way to generate NADPH. In addition, 6PG can also be converted to glycogen or starch and stored. Glucose-6-phosphate dehydrogenase catalyzes the formation of glucose-6-phosphate and NADPH from 6PG and NADP+. NADPH can make WST-8 orange-yellow under the action of 1-mPMS, and the content of 6PG can be calculated by measuring the absorbance value at 450 nm.

GCDH (EC 1.1.1.47) catalyzes D-glucose and NAD(P) to form D-Gluconic acid and NAD(P)H, which are mainly found in the liver of many microorganisms and higher animals. The use of GCDH in the production of oligofructose not only removes glucose from oligofructose and increases its content, but also generates gluconic acid that combines with calcium ions to form calcium gluconate, which is an ideal calcium supplement. Therefore, GCDH has become an ideal enzyme for preparing high content oligofructose. GCDH catalyzes the formation of d-Gluconic acid and NADH from D-glucose and NAD. The activity of glucose dehydrogenase can be reflected by the change of absorbance value of NADH at 340 nm.

Methylcitrate synthase (MCS) exists widely in the mitochondrial matrix of animals, plants, microorganisms and cultured cells, and participates in the regulation of tricarboxylic acid cycle together with citrate synthase (CS). CheKineTM Micro Methylcitrate Synthase (MCS) Activity Assay Kit can be used to detect biological samples such as animal and plant tissues,Cells, Serum orother Liquid samples. In this kit, MCS catalyzes propionyl CoA and oxaloacetic acid to produce methyl citric acid, which further hydrolyzes to methyl citric acid, which promotes the conversion of colorless DTNB to yellow TNB with characteristic absorbance at 412 nm.

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.