QuantiChrom™ β-N-Acetylglucosaminidase Assay Kit
Protocol
SDS
Application
- β-N-Acetylglucosaminidase activity determination in biological samples.
Key Features
- Fast and sensitive. Linear detection range (20 µL sample): 0.2 to 50 U/L for a 30 minute reaction at 37°C.
- High-throughput. Homogeneous "mix-incubate-measure" type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.
Method
- OD405nm
Samples
- Urine, serum, plasma, cell lysate, etc.
Species
- All
Size
- 100 tests
Detection Limit
- 0.2 U/L
Shelf Life
- 12 months
More Details
- β-N-Acetylglucosaminidase (NAG) is a lysosomal enzyme involved in a variety of biological processes such as the degradation of glycoproteins and glycolipids, cell proliferation, and signal transduction. NAG is found in many tissues in the body, but due to its high molecular weight, it can not be filtered through the glomerular membrane. For this reason, in the presence of tubular damage or a glomerular lesion, urinary NAG activity increases. Elevated NAG levels in urine are an early indication of renal damage, such as injury due to diabetes mellitus, inflammation, nephritic syndrome, urinary tract infection, and more. Various forms of cancer have been associated with increased levels of NAG in serum. Genetically inherited lipid storage disorders, such as Tay-Sachs and Sandhoff disease, arise from deficiencies of the enzyme. BioAssay Systems non-radioactive, colorimetric NAG assay is based on the cleavage of p-nitrophenol from a synthetic substrate. p-Nitrophenol becomes intensely colored after addition of the stop reagent. The increase in absorbance at 405 nm after addition of the stop reagent is directly proportional to the enzyme activity.
How do I store the kit?
This kit is shipped at room temperature. Upon receiving, please store the substrate at -20°C and the remainder of the kit in the refrigerator (4°C).How do I prepare cell samples for assays?
Collect cells by centrifugation at 2,000 x g for 5 min at 4°C. For adherent cells, do not harvest cells using proteolytic enzymes; rather use a rubber policeman. Homogenize (10-20 passes in a Dounce homogenizer on ice) or sonicate cells (preferably performed in an ice water bath) in an appropriate volume of cold PBS, approximately one million cells per mL. Centrifuge at 14,000 x g for 10 min at 4°C. Remove supernatant for assay. It is prudent to run a pilot test of the sample at different dilutions. Choose a dilution with the readings in the linear range of the standard curve for further assays. Most samples can be stored at -80°C if not assayed immediately.Do I need to use a standard or standard curve with each assay run?
Yes, it is highly recommended.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.
Zager, R. A., et al. (2021). Iron sucrose ('RBT-3 ') activates the hepatic and renal HAMP1 gene, evoking renal hepcidin loading and resistance to cisplatin nephrotoxicity. Nephrology, Dialysis, Transplantation: Official Publication of the European Dialysis and Transplant Association - European Renal Association 36(3): 465-474. Assay: β-N-Acetylglucosaminidase in mouse plasma.
Johnson, A. C., et al. (2019). Parenterial iron sucrose-induced renal preconditioning: Differential ferritin heavy and light chain expression in plasma, urine, and internal organs. American Journal of Physiology. Renal Physiology 317(6): F1563-F1571. Assay: β-N-Acetylglucosaminidase in human urine.
To find more recent publications, please click here.
Johnson, A. C., et al. (2019). Parenterial iron sucrose-induced renal preconditioning: Differential ferritin heavy and light chain expression in plasma, urine, and internal organs. American Journal of Physiology. Renal Physiology 317(6): F1563-F1571. Assay: β-N-Acetylglucosaminidase in human urine.
To find more recent publications, please click here.
If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.
- Fast turnaround
- Quality data
- Low cost
Please email or call 1-510-782-9988 x 2 to request assay service.
- Fast turnaround
- Quality data
- Low cost
Please email or call 1-510-782-9988 x 2 to request assay service.