There is no guarantee that this assay will detect phosphate covalently bound to a protein. It is possible that the acidic conditions of the working reagent will free the bound phosphate and detect it, but this has not been tested. This assay has a lower limit of detection of 0.4 uM free phosphate.
MAK308
Phosphate Assay Kit
sufficient for 500 colorimetric tests
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.
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usage
sufficient for 500 colorimetric tests
detection method
colorimetric
relevant disease(s)
gastrointestinal diseases; cardiovascular diseases
storage temp.
2-8°C
Related Categories
General description
Phosphate is one of the most abundant minerals in the human body and contains about 1 % of total body weight. Phosphate homeostasis is maintained by the several regulators such as parathyroid hormone (PTH), calcitriol and phosphatonins. Maintenance of phosphate homeostasis is essential for various biological processes, such as energy metabolism, cell signaling, regulation of protein synthesis, skeletal development and bone integrity.[1] An adequate level of phosphate is needed for the process of apoptosis of mature chondrocytes in the growth plate. Phosphate deficiency is associated with the bone pathology and clinical illness.[1]
Application
This kit may be used to measure the liberation of free orthophosphate in the following applications:
- Phosphatase Assays: liberation of phosphate from peptide, protein, or small molecule substrate.
- Lipase Assays: liberation of phosphate from phospholipids
- Nucleoside Triphosphatase Assays: liberation of phosphate from nucleoside triphosphates (ATP, GTP, TTP, CTP etc).
- Quantitation of phosphate in phospholipids, proteins and DNAs, etc.
- Drug Discovery: high-throughput screen for phosphatase inhibitors.
Features and Benefits
Reagent very stable: Due to our innovative formulation, no precipitation of reagent occurs. Therefore no filtration of reagent is needed prior to assays, as is often required with other commercial kits.
- High sensitivity and wide detection range: detection of as little of 20 pmoles of phosphate and useful range between 0.4–50 µM phosphate.
- Fast and convenient: homogeneous “mix-andmeasure” assay allows quantitation of free phosphate within 30 minutes.
- Compatible with routine laboratory and HTS formats: assays can be performed in tubes, cuvettes, or microplates, on spectrophotometers and plate readers.
- Robust and amenable to HTS: Z factors of 0.7–0.9 are observed in 96 well plates. Can be readily automated on HTS liquid handling systems.
Suitability
Suitable for phosphate determination in enzyme reactions with free phosphate releases.
Principle
The Phosphate Assay Kit is based on a proprietary formulation of the malachite green dye. The kit reagent forms a blue colored complex with free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600 - 660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be performed in tubes, cuvettes or multi-well plates. The assays can be conveniently executed in 96-well plates for high-throughput screening of enzyme inhibitors.
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Met. Corr. 1
Storage Class Code
8B - Non-combustible corrosive hazardous materials
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Can this assay be used to measure phosphate from Tyrosine phosphorylated proteins? I have SH2 domain proteins, which binds to pY-proteins. I want to measure the amount of phosphate in each SH2 sample before phosphopeptide enrichment and MS-analysis
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How is shipping temperature determined? And how is it related to the product storage temperature?
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Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf
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How can I determine the shelf life / expiration / retest date of this product?
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If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/449/386/product-dating-information-mk.pdfHelpful?
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What is the recommended method for preparing tissue and cell samples for use with the MAK308-1KT Phosphate Assay Kit? Should detergent or another solution be used for these samples?
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Utilizing mechanical lysis over detergent-mediated lysis whenever feasible. This is particularly crucial in this instance as detergents could lead to undesired phosphate contamination. They suggest employing a Dounce Homogenizer or sonication. Additionally, the extent of tissue and/or cell lysis can be visually verified using a microscope, if accessible.
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