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MAK307

Sigma-Aldrich

Malachite Green Phosphate Assay Kit

sufficient for 2500 colorimetric tests

Synonym(s):

Phosphate Determination Assay Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

usage

sufficient for 2,500 colorimetric tests

detection method

colorimetric

relevant disease(s)

cardiovascular diseases; gastrointestinal 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.† 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.†

Application

Malachite Green Phosphate Assay Kit has been used in phosphate quantification[1] and steady-state kinetics.[2]
Malachite Green Phosphate Assay Kit has been used to measure the liberation of free orthophosphate in the following applications:
  • Phosphatase Assays: liberation of phosphate from peptide, protein, or small molecule substrate.[1][3]
  • 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

Compatible with high-throughput handling systems.

Suitability

Suitable for phosphate determination in a variety of biological and environmental samples.

Principle

Malachite Green is a basic dye belonging to the triphenylmethane group. It is used in biology for the staining of microscopic samples, such as plant tissue, yeast and bacteria. The malachite green method is a sensitive method for the detection of released phosphate.†† The Malachite Green Phosphate Assay Kit is based on quantification of the green complex formed between Malachite Green, molybdate and 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 executed in tubes, cuvettes or multi-well plates. The assays can be conveniently performed in 96 and 384 well plates for high throughput screening of enzyme inhibitors.

Pictograms

FlameCorrosion
GHS02,GHS05

Signal Word

Warning

Hazard Statements

H226,H290,H315,H319

Hazard Classifications

Eye Irrit. 2 - Flam. Liq. 3 - Met. Corr. 1 - Skin Irrit. 2

Storage Class Code

3 - Flammable liquids

Flash Point(F)

82.4 °F

Flash Point(C)

28 °C

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
307CE08A20
307CE08A13
307CE03A11
307CD12A06
307CD08A28

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If you require a particular version, you can look up a specific certificate by the Lot or Batch number.

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Edward G.
Synthetic Dyes in Biology, Medicine And Chemistry (2012)
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Human molecular genetics, 30(3-4), 198-212 (2021-02-01)
Lowe Syndrome (LS) is a lethal genetic disorder caused by mutations in the OCRL1 gene which encodes the lipid 5' phosphatase Ocrl1. Patients exhibit a characteristic triad of symptoms including eye, brain and kidney abnormalities with renal failure as the
On targeted proteins involved in the neuroprotective effects of lithium citrate.
Torshin I Y, et al.
Neurology, neuropsychiatry, psychosomatics, 9(1), 2017-2017 null
Ilke Sen et al.
Nature communications, 11(1), 138-138 (2020-01-11)
In C. elegans, the conserved transcription factor DAF-16/FOXO is a powerful aging regulator, relaying dire conditions into expression of stress resistance and longevity promoting genes. For some of these functions, including low insulin/IGF signaling (IIS), DAF-16 depends on the protein SMK-1/SMEK
Methods in Plant Cell Biology, 50 (1995)
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Questions

1–9 of 9 Questions  

  1. How can I determine the shelf life / expiration / retest date of this product?

    1 answer

    1. 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.pdf

      Helpful?

  2. Is this kit able to detect pyrophosphate too?

    1 answer

    1. This kit is intended for the detection of free orthophosphate and has not been tested for pyrophosphate detection. There are other products available that are specifically for use in pyrophosphate detection. Please see the links below to review each item:
      MAK168, Pyrophosphate Assay Kit
      https://www.sigmaaldrich.com/product/sigma/mak168
      MAK169. High Sensitivity Pyrophosphate Assay Kit
      https://www.sigmaaldrich.com/product/sigma/mak169

      Helpful?

  3. What are the reasons behind the suggestion to use lower concentrations than 0.25mM? Are there potential unwanted biochemical reactions between GTP and malachite green/molybdate if GTP concentration exceeds 0.25mM?

    1 answer

    1. The kit operates by releasing free orthophosphate from ATP or GTP, followed by the creation of a green complex involving the orthophosphate, molybdate, and Malachite green. It has been observed that samples containing ATP and GTP at concentrations exceeding 0.25mM result in the measurement of free phosphate levels falling outside the detection range of the kit (0.02 µM and 40 µM phosphate), and hence should be avoided.

      Helpful?

  4. Does the product MAK307 contain magnesium?

    1 answer

    1. Magnesium is not included as a component in any of the reagents provided with the kit for product MAK307. If magnesium serves as an essential cofactor for the enzyme, it is recommended to supplement it in the buffer before conducting the phosphate determination assay.

      Helpful?

  5. Why is it necessary to dilute the ATP concentration to <0.25 mM before the assay? Does ATP interfere with the assay?

    1 answer

    1. BioAssay Support recommends reducing the ATP concentration before the assay to minimize background interference resulting from non-enzymatic ATP hydrolysis and to decrease the likelihood of reagent precipitation with unhydrolyzed ATP.

      Helpful?

  6. our kit expired, when I used to measure extracted polyphosphate degraded by the enzyme, the results were so different compared to the precious ones (when the kit was not expired), this means the result was not reliable after it expired?

    1 answer

    1. Correct, products used beyond the expiration date will likely offer poor results.

      Helpful?

  7. Does Malachite Green interact with glycerol?

    1 answer

    1. The Malachite Green Phosphate Assay Kit is based on the quantification of the green complex formed between Malachite Green, molybdate, and free orthophosphate. No specific compatibility study has been done with this kit and glycerol. This would need to be tested empirically, in a sample with a known phosphate concentration, with and without glycerol at the desired concentration.

      Helpful?

  8. Can we test the same MG in shrimp matrix ?

    1 answer

    1. This product is suitable for a variety of sample types. There is no reason to suspect a problem with shrimp matrix. Attention should be given to sample preparation, handling, and dilution. See the link below to review the product datasheet which includes the protocol and troubleshooting tips:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/171/212/mak307bul.pdf

      Helpful?

  9. How is shipping temperature determined? And how is it related to the product storage temperature?

    0 answers

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