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#DIPI-500 QuantiChrom™ Phosphate Assay Kit
Product name : QuantiChrom™ Phosphate Assay Kit
Catalog number : DIPI-500
Supplier name : Bioassays
Data sheet: Ask more or other datasheet now !
About this Product :QuantiChrom™ Phosphate Assay Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ Phosphate Assay Kit.
More Details about
QuantiChromTM Phosphate Assay Kit (DIPI-500)
Quantitative Colorimetric Phosphate Determination at 620nm
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. BioAssay Systems' 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.
- 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.
- 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.
KIT CONTENTS (500 tests in 96-well plates)
Reagent: 50 mL
Pi standard: 14 mL 0.28 mg/dL (30 μM)
Blank Control: 14 mL
Storage conditions. The kit is shipped at room temperature. Store all components at 4°C. Shelf life: 12 months after receipt.
Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents.
Please refer to Material Safety Data Sheet for detailed information.
Important: bring reagents to room temperature and shake before use.
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.
The phosphate concentration of Sample is calculated as
= (ODSAMPLE – ODBLANK) / (ODSTANDARD – ODBLANK) X 0.28 (mg/dL)
ODBLANK, ODSTANDARD and ODSAMPLE are OD620nm values of Blank, Standard and Sample, respectively.
Conversions: 1 mg/dL Pi equals 105.3 μM, 0.001% or 10 ppm.
MATERIALS REQUIRED, BUT NOT PROVIDED
Pipeting devices and accessories.
Procedure using 96-well plate:
Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Spectrophotometer and cuvets for measuring OD 620nm.
EXAMPLES (96-well plate assay):
1. 6.4 ± 0.6
2. 2.2 ± 0.1
3. 3.5 ± 0.1
4. 0.081 ± 0.003
5. 0.003 ± 0.001
6. 0.02 ± 0.001
7. 1.10 ± 0.01
8. 0.56 ± 0.06
9. 0.19 ± 0.03
Biological Samples: 1. Commercial 2% reduced fat milk (Kirkland). 2. Invitrogen fetal bovine serum. 3. Fresh human urine. Water samples: 4. Tap water (Hayward, CA). 5. Tap water (San Bruno, CA). Food and Beverages: 6. Crystal Geyser natural alpine spring water. 7. Coca-cola® classic coke. 8.
Lipton Lemon iced tea. Environmental: 9. Soil extract. 5.6 g of soil (Hayward, CA) was extracted with 10 mL MilliQ water. The supernatant was centrifuged to remove any insoluble particles. Clear supernatant was assayed.
1. Abranches, J. (2008). CcpA regulates central metabolism and virulence gene expression in Streptococcus mutans. J Bacteriol. 190(7):2340-9.
2. Hildebrand, J. et al (2009) Functional and energetic characterization of P-gp-mediated doxorubicin transport in rainbow trout. Comp Biochem Physiol C Toxicol Pharmacol. 149(1):65-72.
3. Dunbar, D.R. et al. (2010). Transcriptional and physiological responses to chronic ACTH treatment by the mouse kidney. Physiol Genomics 40(3): 158-166.
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WP1004: Kit Receptor Signaling Pathway
WP1028: Pentose Phosphate Pathway
WP1121: Kit Receptor Signaling Pathway
WP1147: Pentose Phosphate Pathway
WP122: Pentose Phosphate Pathway
WP1231: Pentose Phosphate Pathway
WP134: Pentose Phosphate Pathway
WP1341: Kit Receptor Signaling Pathway
WP147: Kit Receptor Signaling Pathway
WP1493: Carbon assimilation C4 pathway
WP1657: Glycerolipid metabolism
WP1658: Glycerophospholipid metabolism
WP1664: Inositol phosphate metabolism
WP1684: Pentose phosphate pathway
WP2185: Purine metabolism
WP220: Ribose and Deoxyribose Phosphate Metabolism
WP2212: Methylerythritol phosphate pathway
WP228: Deoxyribose phosphate metabolism
WP2340: Thiamine (vitamin B1) biosynthesis and salvage
WP2341: vitamin B1 (thiamin) biosynthesis and salvage pathway
WP260: Glucose-1-phosphate metabolism
WP282: Pentose Phosphate Pathway
WP312: Pentose Phosphate Pathway
WP369: Pentose Phosphate Pathway
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