| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Biological (e.g. serum) samples |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitative determination of acid phosphatase activity in biological samples. The assay uses FL360/450 for signal readout. Compatible sample input includes Biological (e.g. serum) samples. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: FL360/450 supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Biological (e.g. serum) samples, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.008 U/L for interpreting low-signal samples.
- Feature emphasis: Fast and sensitive. Linear detection range (20 µL sample): 0.008 to 10 U/L for a 30-minute reaction.
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 acid 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
Acid Phosphatase(ACP) is an enzyme that catalyzes the cleavage of phosphate groups from other molecules during digestion. Acid phosphatase can be found in lysosomes and become active after fusing with endosomes, acidifying the pH and, thus, creating an optimal environment for ACP. ACP can also be found in bone, spleen, liver, kidney, and blood. Serum levels can be used as a biomarker for prostatic carcinoma, although prostate-specific antigen (PSA) is more widely used.BioAssay System’s non-radioactive, fluorimetric ACP assay is based on the cleavage of methylumbelliferyl phosphate. Methylumbelliferone becomes intensely fluorescent after the addition of the stop reagent. The increase in fluorescence at 360/450 nm after the addition of the stop reagent is directly proportional to the enzyme activity.
Detection method
Fluorescent (FL 360/450 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.008 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.
- Short assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.
Common research applications
- Quantify acid phosphatase in biological (serum) by FL360/450 readout.
- Compare treatment or phenotype groups using matched biological (serum) handling.
- Monitor time-course or pre/post changes in biological (serum) 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.
I don’t have the correct wavelength filter, will the assay still work?
Typically if you are within ± 20 nanometers the assay will still work, but have slightly reduced sensitivity. Some assays have a narrower peak than others, call our tech support for a spectrum to verify.
I don’t have a 96-well plate, will the assay kit work in a 384 well plate, cuvette, etc?
Yes, the assay can be used in any standard plate and cuvette. Simply adjust the total reaction volume to your container. For 384 well use 50 μL, for a cuvette use 500 μL or 1 mL.
Does the assay kit work in particular species?
Yes, this assay works in all species.
How do I prepare cell or tissue samples for assays?
Please refer to the protocol for sample preparation:
Blood samples: Serum and plasma should be diluted 2-5 fold.
Blood samples
Tissue: Prior to dissection, rinse tissue in Tris buffered saline (pH 7.4) to remove blood. Homogenize tissue (50 mg) in 200 μL 50 mM Tris buffer (pH 7.5). Centrifuge at 14,000 x g for 10 min at 4°C. Remove supernatant for assay.
Tissue
Cell lysate: 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 or sonicate cells in an appropriate volume of cold 50 mM Tris buffer (pH 7.5), approximately one million cells per mL. Centrifuge at 14,000 x g for 10 min at 4°C. Remove supernatant for assay. All samples can be stored at -80 to -20°C for at least one month.
Can I store unused reagents for future use?
Yes, unused reagents can be stored according to the assay protocol. Repeated freeze/thaw cycles of reagents should be avoided. Since Assay Buffer is very light sensitive, always keeps buffer in the dark (amber bottle or tube) as much as possible for long term usage.
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.
