Product Details
| abID | ab65622 |
|---|---|
| Cat # +Size | K410-500 |
| Size | 500 assays |
| Detection Method | Absorbance (650 nm) |
| Species Reactivity | Mammalian |
| Applications | Phosphate concentrations between 1 µM and 1 mM, with a lower limit of detection of approximately 0.1 nmol, can be directly determined. |
| Features & Benefits | • Simple procedure; takes ~ 3 hours • Fast and convenient • The assay is sensitive and stable • The TUNEL-based assay kit provides complete components including positive and negative control cells for convenient detection of DNA fragmentation in cultured cells and tissue sections. |
| Kit Components | • Phosphate Reagent • Phosphate Standard (10 mM) |
| Storage Conditions | RT |
| Shipping Conditions | RT |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Phosphate is one of the most important of the inorganic ions in biological systems. It functions in a variety of roles. One of the most important roles is as a molecular switch, turning enzyme activity on and off through the mediation of the various protein kinases and phosphatases in biological systems. Phosphate is also of great importance in mineralization processes and is a primary stimulus of algal blooms frequently found in bodies of fresh water, due to run-off from areas of high fertilizer use. The newly designed Phosphate Colorimetric Assay Kit provides an easy, quick and sensitive means of assessing phosphate over a wide range of concentrations. The assay utilizes a proprietary formulation of malachite green and ammonium molybdate which forms a chromogenic complex with phosphate ion giving an intense absorption band around 650 nm. Phosphate concentrations between 1 µM and 1 mM, with a lower limit of detection of approximately 0.1 nmol, can be directly determined. The Phosphate Colorimetric Assay Kit provides 500 assays using microtiter plates or 100 assays using 1 ml cuvettes.
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Can K410 Phosphate Colorimetric Assay Kit be applied in samples with high citrate concentration? Is molybdate reagent solution required for this kit? Could you help to check and suggest?
K410 is based on the reaction of malachite green and ammonium molybdate with phosphate ions, which forms a chromogenic complex giving an intense absorption band around 650 nm. Molybdate reagent is already added in one of the components and it is required for the assay. Citrate may be inhibitory as it may form complexes with free molybdate and prevent any further color development. However, the titration range for citrate that may be inhibitory has not been tested.
What are the possible sources of interferences for this assay?
1. We ask our customers to avoid using glasswares since a lot of laboratory detergents contain high amounts of phosphate. Disposable plastic wares are therefore recommended for the assay and sample preparation.
2. Presence of IgG and IgM paraproteins can produce false increase in Phosphate concentration by producing precipitates in the reaction. Dilution of the sample can reduce the interference. Deproteination of the samples works best (Clin Chem. 1995 Apr;41(4):60914.).
3. It's advisable to avoid TritonX during sample preparation
4. Some reports suggests Trichloroacetic acid and Tungtic acid can interfere with the assay
5. EDTA, oxalate and citrate should be avoided
6. Significant interference can be caused by hemolysate, bilirubin and intralipids.
2. Presence of IgG and IgM paraproteins can produce false increase in Phosphate concentration by producing precipitates in the reaction. Dilution of the sample can reduce the interference. Deproteination of the samples works best (Clin Chem. 1995 Apr;41(4):60914.).
3. It's advisable to avoid TritonX during sample preparation
4. Some reports suggests Trichloroacetic acid and Tungtic acid can interfere with the assay
5. EDTA, oxalate and citrate should be avoided
6. Significant interference can be caused by hemolysate, bilirubin and intralipids.
Do you use a stabilizer in the Phosphate Reagent (K410-500-1) provided with the kit?
Yes, we use a stabilizer for phosphate in the Phosphate Reagent provided with the kit.
Chen et al., PDGFB-based stem cell gene therapy increases bone strength in the mouse. PNAS, Jul 2015; 112: E3893 - E3900.
Krieger et al., Effect of Potassium Citrate on Calcium Phosphate Stones in a Model of Hypercalciuria. J. Am. Soc. Nephrol., Apr 2015; 10.1681/ASN.2014121223.
Burris et al., Estrogen directly and specifically downregulates NaPi-IIa through the activation of both estrogen receptor isoforms (ERα and ERβ) in rat kidney proximal tubule. Am J Physiol Renal Physiol, Mar 2015; 308: F522 - F534.
Prazeres et al., Ocean acidification induces biochemical and morphological changes in the calcification process of large benthic foraminifera. Proc R Soc B, Mar 2015; 282: 20142782.
Wu-Wong et al., Vitamin D receptor agonist VS-105 improves cardiac function in the presence of enalapril in 5/6 nephrectomized rats. Am J Physiol Renal Physiol, Feb 2015; 308: F309 - F319.
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