QuantiFluo™ Alkaline Phosphatase Assay Kit

Overview

For quantitative determination of alkaline phosphatase (ALP) activity using stable 4-methylumbelliferyl phosphate substrate. The assay uses FL360/450nm for signal readout. Compatible sample input includes Serum, plasma etc. Typical stated assay timing is 20 min.

Key elements and design rationale

• Readout format: FL360/450nm supports plate-based signal acquisition and consistent comparison across matched samples.
• Sample compatibility: The stated sample scope includes Serum, plasma 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 0.02 U/L for interpreting low-signal samples.
• Feature emphasis: High sensitivity and wide linear range. Use 10 µL sample. The detection limit of 0.02 U/L (20 min reaction).

Additional feature notes highlight Homogeneous and simple procedure. A simple “mix-and-measure” procedure allows reliable quantitation of ALP activity within 20 minutes; Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments. Available format information for this listing includes 100 Tests.

Biological background

This product is centered on measurement of alkaline phosphatase 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

Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in an alkaline environment, resulting in the formation of an organic radical and inorganic phosphate. In mammals, this enzyme is found mainly in the liver and bones. Marked increase in serum ALP levels, a disease known as hyperalkalinephosphatasemia, has been associated with malignant biliary obstruction, primary biliary cirrhosis, primary sclerosing cholangitis, hepatic lymphoma and sarcoidosis. Simple, direct and automation-ready procedures for measuring ALP activity in serum are becoming popular in Research and Drug Discovery. This improved method utilizes 4-methylumbelliferyl phosphate that is hydrolyzed by ALP into a highly fluorescent product 4-methylumbelliferone. The rate of the fluorescence increase is directly proportional to the enzyme activity.

Detection method

Fluorescent (FL 360/450 nm).

Detection limit and analytical sensitivity

Reported detection limit: 0.02 U/L.

Procedures and timing

Stated procedure or timing information: 20 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.
• Short assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.

Common research applications

• Quantify alkaline phosphatase in serum, plasma by FL360/450 nm readout.
• Compare treatment or phenotype groups using matched serum, plasma handling.
• Monitor time-course or pre/post changes in serum, plasma 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.

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.