Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Tissue, Suspension cells, Serum, Adherent cells,
Assay type:Enzyme activity,
Sensitivity:= 50 µU,
Assay time:25m,
Assay Platform:Microplate reader
Product details:
N-Acetylglucosaminidase (beta-NAG) Activity Assay Kit (Colorimetric) (ab204705) provides a simple and sensitive method for monitoring NAG enzymatic activity.
How the assay works
In the N-Acetylglucosaminidase (beta-NAG) assay, NAG uses a synthetic substrate, like p-nitrophenyl-N-acetyl-beta-D-glucosaminide (pNP-GlcNAc), to release a detectable product, p-nitrophenol (pNP), which can be measured by absorbance (400 nm).
N-Acetylglucosaminidase (beta-NAG) assay protocol summary
- Add samples and standards to wells
- Add NAG Substrate to sample wells and incubate for 5-30 minutes
- Add Stop Solution and incubate for 10 minutes
- Analyze with a microplate reader to measure optical density (OD 400 nm).
How other researchers are using ab204705
N-Acetylglucosaminidase (beta-NAG) Activity Assay Kit (Colorimetric) has been used with a variety of sample types including:
Human iPSC-derived lacrimal glands
Calf adipocytes
Blood and urine sample from diabetic mice
References: 1-Asal M et al. 2023, 2-Yu H et al. 2023, 3-de Groot T et al. 2023
Other Notes
This product was previously called K733 Biovision Beta-N-Acetylglucosaminidase Activity Assay Kit (Colorimetric). Biovision was acquired by Abcam in 2021.
Beta-N-Acetylglucosaminidase (NAG, EC 3.2.1.52) is a lysosomal enzyme that is expressed in various tissues, including kidney, liver and lungs. NAG can cleave N-acetyl-glucosamine, a monosaccharide derivative of glucose. Its concentration in urine is minimal due to its inability to cross the glomerular basal membrane. Increased concentration of NAG in urine indicates renal tubular cell breakdown. Acute Kidney Injury (AKI) is the sudden loss of kidney functions, causing electrolyte imbalance, and retention of urea and other nitrogenous products. NAG has become one of the most studied and used biomarkers for the detection and diagnosis of AKI.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
N-Acetylglucosaminidase also known as NAG β-NAG and NAG-5 plays an important role in the breakdown of glycoproteins by catalyzing the hydrolysis of terminal N-acetylglucosamine residues in N-acetyl-β-D-glucosaminides. This enzyme has a molecular mass of approximately 55 kDa. It expresses in various tissues with noticeable activity in lysosomes and is critical for maintaining cellular homeostasis by processing glycosylated proteins.
Biological function summary
The enzyme N-acetylglucosaminidase is essential for cell growth signaling and structural integrity. It belongs to a larger group known as glycoside hydrolases which cooperate during the degradation of complex carbohydrates and glycoproteins. NAG's function impacts the lysosomal degradation pathway where it often works with substrates like glycosaminoglycans ensuring the recycling of cellular components in the lysosomal environment.
Pathways
N-acetylglucosaminidase integrates with lysosomal storage pathways and takes part in glycosaminoglycan degradation. Its activity is keenly tied with proteins such as β-glucuronidase working alongside in the sequential breakdown of glycan chains. These pathways are important for cellular waste disposal and recycling contributing to overall metabolic harmony in cells.
N-acetylglucosaminidase links to conditions such as mucopolysaccharidosis and Tay-Sachs disease. Deficiency in NAG enzymatic function leads to improper degradation of glycosaminoglycans or gangliosides contributing to their pathophysiology. The enzyme shares this relation with proteins like Hexosaminidase A in Tay-Sachs disease demonstrating its pivotal role in neurological health.
