Phosphate Assay Kit

Catalog No. ABIN1000269

Product Details

Target: Phosphate

Application: Biochemical Assay (BCA)

Sample Type: Beverages, Cell Culture Supernatant, Fertilizer, Food, Saliva, Serum, Soil, Sweat, Urine, Water

Specificity: 3 μg/dL (0.3 μM)

Characteristics: Sensitive and accurate. Linear detection range 0.30 µM (0.0028 mg/dL) to 50 µM (0.47 mg/dL) phosphate in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 30 min. Can be readily automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Assays can be executed in cuvet or 96-well plate.
Low interference in biological samples.
No pretreatments are needed. Assays can be directly performed on raw biological samples i.e., in the presence of lipid, protein and minerals.

Components: Reagent: 50 mL. Pi standard: 14 mL 0.28 mg/dL (30 µM). Blank Control: 14 mL.

Material not included: Pipeting devices and accessories. Clear bottom 96-well plates (e.g. Corning Costar) and plate reader. Spectrophotometer and cuvets for measuring OD 620nm.

Application Details

Application Notes: Direct Assays: Pi in serum, urine, saliva, sweat, tissue culture etc.
Drug Discovery/Pharmacology: effects of drugs on Pi metabolism.
Food and Beverages: Pi determination.
Environment: Pi determination in water, soil and fertilizer.

Protocol: Procedure using 96-well plate:
1. Set up standards and samples. Transfer 50 µL distilled water (Blank), Standard and samples in duplicate wells of a clear bottom 96-well plate.
2. Add 100 µL Reagent and tap lightly to mix.
3. Incubate 30 min at room temperature and read optical density at 620nm (600-660nm).

Procedure using cuvette:
1. Set up test tubes labeled Blank, Standard, Samples. Transfer 400 µL Water, Standard and samples to appropriately labeled tubes.
2. Add 800 µL Reagent and tap lightly to mix.
3. Incubate 30 min at room temperature, transfer to cuvet and read optical density at 620 nm (600-660nm). Important: (1) if sample OD is higher than the OD for standard, dilute samples in distilled water and repeat the assay. (2) It is not necessary to prepare a calibration curve, because the concentration of the provided standard lies within the linear range. (3) Precipitation may occur at high concentrations of phosphate (>100 µM), or in the presence of high concentrations of e.g. proteins and metals. In this case, dilute samples in distilled water and repeat the assay.

Reagent Preparation:

Important: bring reagents to room temperature and shake before use.

Calculation of Results:

Conversions: 1 mg/dL Pi equals 105.3 µM, 0.001% or 10 ppm.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Dunbar, Khaled, Evans, Al-Dujaili, Mullins, Mullins, Kenyon, Bailey: "Transcriptional and physiological responses to chronic ACTH treatment by the mouse kidney." in: Physiological genomics, Vol. 40, Issue 3, pp. 158-66, (2010) (PubMed).

Ponda, Barash, Feig, Fisher, Skolnik: "Moderate kidney disease inhibits atherosclerosis regression." in: Atherosclerosis, Vol. 210, Issue 1, pp. 57-62, (2010) (PubMed).

Galitzer, Ben-Dov, Silver, Naveh-Many: "Parathyroid cell resistance to fibroblast growth factor 23 in secondary hyperparathyroidism of chronic kidney disease." in: Kidney international, Vol. 77, Issue 3, pp. 211-8, (2010) (PubMed).

OConnor, Zayzafoon, Farach-Carson, Schanen: "Mecp2 deficiency decreases bone formation and reduces bone volume in a rodent model of Rett syndrome." in: Bone, Vol. 45, Issue 2, pp. 346-56, (2009) (PubMed).

Xie, Knight, Leggett: "Comparison of media used to evaluate Rhizobium leguminosarum bivar viciae for phosphate-solubilizing ability." in: Canadian journal of microbiology, Vol. 55, Issue 7, pp. 910-5, (2009) (PubMed).

Abranches, Nascimento, Zeng, Browngardt, Wen, Rivera, Burne: "CcpA regulates central metabolism and virulence gene expression in Streptococcus mutans." in: Journal of bacteriology, Vol. 190, Issue 7, pp. 2340-9, (2008) (PubMed).

Hildebrand, Bains, Lee, Kennedy: "Functional and energetic characterization of P-gp-mediated doxorubicin transport in rainbow trout (Oncorhynchus mykiss) hepatocytes." in: Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, Vol. 149, Issue 1, pp. 65-72, (2008) (PubMed).

Jubeck, Gohr, Fahey, Muth, Matthews, Mattson, Hirschmugl, Rosenthal: "Promotion of articular cartilage matrix vesicle mineralization by type I collagen." in: Arthritis and rheumatism, Vol. 58, Issue 9, pp. 2809-17, (2008) (PubMed).

Chung, Ahn, Jeon, Lee, Lee, Tae: "Enhanced bone regeneration with BMP-2 loaded functional nanoparticle-hydrogel complex." in: Journal of controlled release : official journal of the Controlled Release Society, Vol. 121, Issue 1-2, pp. 91-9, (2007) (PubMed).

Target Details

Target: Phosphate

Background: Quantitative determination of phosphate by colorimetric (620nm) method.
Procedure: 30 min.

Phosphate (Pi) is one of the most important ion species in nature. Phosphate is present in all biological systems. It is a major constituent in minerals and fertilizers, and is a component of industrial wastewater. Thus accurate determination of phosphate concentration finds numerous applications in pharmacology, biomedical research, clinical chemistry, industrial process monitoring and environmental monitoring. Simple, direct and automation-ready procedures for measuring phosphate concentration in biological and environmental samples are becoming popular. This phosphate assay kit is designed to measure phosphate ion directly in samples without any pretreatment. The improved Malachite Green method utilizes the malachite green dye and molybdate, which forms a stable colored complex specifically with inorganic phosphate. The intensity of the color, measured at 620nm, is directly proportional to the phosphate concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples.