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Cat. No.: NAGAK-100 (for 100T)

Description

N-Acetylglucosaminidase Activity Assay Kit, Colorimetric, abbreviated as NAG Assay Kit, is a colorimetric assay kit designed for the rapid and highly sensitive detection of N-acetylglucosaminidase activity in biological fluids such as serum, plasma, and urine, as well as in tissue and cell lysates, and cell culture supernatants.

N-Acetylglucosaminidase (NAG) is an acidic glycosidase located in the lysosomes of cells. It can hydrolyze β-N-acetylglucosaminide and β-N-acetylgalactosaminide and is widely present in various tissues, organs, body fluids, and blood cells, with particularly high concentrations in renal tubules and urinary tract epithelial cells. Due to its large molecular weight, NAG in serum cannot normally pass through the glomerular filter, resulting in very low levels in urine. When kidney damage occurs, the excretion of NAG in urine increases, making urinary NAG measurement a sensitive indicator for detecting kidney injury, especially ischemia and necrosis of renal tubules. Elevated urinary NAG is commonly seen in conditions such as glomerulonephritis, tubulointerstitial lesions, congenital tubular disorders, renal transplant rejection, and lupus nephritis. In cases of acute renal failure caused by shock, urinary NAG enzyme levels can be up to 1200 times the normal value. During rejection reactions following kidney transplantation, this enzyme can appear up to three weeks before conventional renal function tests confirm rejection. The activity detection of N-acetylglucosaminidase is of significant importance for the diagnosis, prevention, and treatment of related clinical diseases.

Components

• Buffer A for Metabolic Assay
• NAG Detection Buffer
• Substrate
• Stop Solution
• p-Nitrophenol Standard Solution (50mM)

Principle

Under acidic conditions, N-acetylglucosaminidase hydrolyzes the substrate p-nitrophenyl-N-acetylglucosaminide to produce free p-nitrophenol. In alkaline conditions, p-nitrophenol appears yellow and has a maximum absorption peak around 400 nm. The amount of yellow product generated in the reaction system is proportional to the activity of N-acetylglucosaminidase in the sample.

Features

• This assay kit has high sensitivity, a wide linear range, and requires a small sample volume. It can detect enzyme concentrations as low as 13 µU with a sample volume of 20 µL and shows a good linear relationship within the activity range of 0.67-66.7 U/L. The kit provides a standard solution of p-nitrophenol, the product of N-acetylglucosaminidase activity, which can be used to establish a standard curve to calculate the activity of N-acetylglucosaminidase in the samples.

• The detection lysis buffer provided in this kit has a certain degree of versatility. Cell or tissue samples lysed using Buffer A for Metabolic Assay from this kit can also be used for sample detection in other metabolic assay kits that use the same Buffer A for Metabolic Assay, demonstrating strong versatility. Additionally, it can be used for protein concentration detection, SDS-PAGE, or Western blotting of easily soluble proteins.

• This kit offers fast detection and a wide range of applications. It can be used to detect biological fluids such as serum, plasma, and urine from mice, rats, and humans, as well as cell culture supernatants and tissue or cell lysates. The entire process can be completed in approximately 0.5-1 hour. This kit is suitable not only for the detection of small sample quantities but also for high-throughput screening in automated systems.

Storage

Store at -20ºC, valid for one year. The substrate and p-nitrophenol standard solution should be protected from light. Avoid repeated freeze-thaw cycles of all reagents.

Precautions

• Buffer A for Metabolic Assay, NAG detection buffer (hereafter referred to as detection buffer), and stop solution should be completely thawed and brought to room temperature before use; otherwise, the detection results may be affected. Other reagents should be used on ice.
• Serum samples stored at 4ºC should not be kept for more than two weeks, as this may affect the accuracy of the detection results. Serum samples are usually stored at -20ºC, with -80ºC being preferable.
• This product is intended for scientific research by professionals only and is not for clinical diagnosis or treatment, food, or drug use, and should not be stored in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves during operation.