| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Urine, serum, plasma, cell lysate, etc |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitatve determination of β-N-Acetylglucosaminidase activity and evaluation of drug modulators. The assay uses OD405nm for signal readout. Compatible sample input includes Urine, serum, plasma, cell lysate, etc. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD405nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Urine, serum, plasma, cell lysate, etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.2 U/L for interpreting low-signal samples.
- Feature emphasis: Fast and sensitive. Linear detection range (20 µL sample): 0.2 to 50 U/L for a 30-minute reaction at 37°C.
Additional feature notes highlight High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of β-n-acetylglucosaminidase within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
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 System’s non-radioactive, colorimetric NAG assay is based on the cleavage of p-nitrophenol from a synthetic substrate. p-Nitrophenol becomes intensely colored after the addition of the stop reagent. The increase in absorbance at 405 nm after the addition of the stop reagent is directly proportional to the enzyme activity.
Detection method
Colorimetric (OD 405 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.2 U/L.
Procedures and timing
Stated procedure or timing information: 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify β-n-acetylglucosaminidase in urine, serum, plasma, cell lysate by OD405 nm readout.
- Compare treatment or phenotype groups using matched urine, serum, plasma, cell lysate handling.
- Monitor time-course or pre/post changes in urine, serum, plasma, cell lysate across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
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 laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Iron sucrose (‘RBT-3 ‘) activates the hepatic and renal HAMP1 gene, evoking renal hepcidin loading and resistance to cisplatin nephrotoxicity
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.
Parenterial iron sucrose-induced renal preconditioning: Differential ferritin heavy and light chain expression in plasma, urine, and internal organs
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.
