Phosphorus (Pi) Colorimetric Assay Kit (Phospho molybdate method)

    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience
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    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K245

      Size:
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $120

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry

      Valid period: 6 months

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      Application

      This kit can be used to measure Phosphorus (Pi) content in serum, plasma, tissue and other samples.

       

      Detection significance

      Phosphorus is an important mineral that maintains cellular energy, mineralizes bones, and protects non-bone tissue from calcification. Inorganic phosphorus is the component of DNA, RNA, ATP and phospholipid. Phosphorus is existed in the form of ester and phosphate anion in the whole blood. The concentration of phosphorus is strictly regulated by specific ion transport proteins and hormones.

       

      Detection principle

      Inorganic phosphorus react with molybdic acid to produce phosphomolybdic acid. Phosphomolybdic acid can be reduced to molybdenum blue under the action of reducing agent. And the molybdenum blue have a maximum absorption peak at 660 nm. The phosphorus content can be calculated indirectly be measuring the OD value at 660 nm.


      Experimental instrument

      Test tube, Micropipettor, Water bath, Vortex mixer, Microplate reader (660 nm)

      Sample preparation

      1. Serum sample:

      Fresh blood was collected and placed at 25 for 30 min to clot the blood. Centrifuge the sample at 4 for 15 min at 2000 ×g, the upper yellowish clear liquid was taken as serum. Place the serum on ice for detection. If not detected on the same day, stored the serum at-80, which can be stored for a month.

       

      2. Plasma sample:

      The fresh blood was added into the test tube containing anticoagulant and mixed upside down. Centrifuge the sample at 4 for 10 min at 700~1000 ×g, the upper yellowish transparent liquid was taken as the plasma, and the middle white interference layer (white blood cells and platelets) could not be absorbed. Place the plasma on ice for detection. If not detected on the same day, stored the serum at-80, which can be stored for a month.

       

      3. 10% tissue homogenate sample:

      Accurately weigh the tissue sample, add 9 times the volume of PBS (0.01 M, pH7~7.4) according to the ratio of Weight (g): Volume (mL) =1:9. Mechanical homogenate the sample in ice water bath. Centrifuge at 10000 g for 10 min, then take the supernatant and preserve it on ice for detection. Meanwhile, determine the protein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165).

      Operation steps

      1. The preparation of sample supernatant

      Take 0.1 mL serum (plasma) or 10% tissue homogenate sample, then add 0.4 mL reagent 4, mix fully. Centrifuge at 1100 × g for 10 min and take the supernatant for detection.

       

      2. The measurement of samples

      1) Blank tube: Take 0.2 mL of double distilled water to an EP tube.

      Standard tube: Take 0.2 mL of 0.5 mmol/L standard solution to an EP tube.

      Sample tube: Take 0.2 mL of sample supernatant to the EP tubes.

      2) Add 2.0 mL of Chromogenic agent into each tube of Step 1 and mix fully.

      3) Incubate the tubes at 37℃ for 30 min, then cool the tubes to room temperature.

      4) Set the spectrophotometer to zero with double distilled water, and measure the OD at 660 nm with 1 cm optical path quartz cuvette.

        

      Note: The following operating table could be as a reference.


       

      Blank tube

      Standard tube

      Sample tube

      Double distilled water (mL)

      0.2

       

       

      0.5 mmol/L standard solution (mL)

       

      0.2

       

      Sample supernatant (mL)

       

       

      0.2

      Chromogenic agent (mL)

      2.0

      2.0

      2.0

      Mix fully, then incubate the tubes at 37 for 30 min, then cool the tubes to room temperature. Set the spectrophotometer to zero with double distilled water, and measure the OD at 660 nm with 1 cm optical path quartz cuvette.


      Technical parameter

      1. The sensitivity of the kit is 0.005-2.0 mmol/L.

      2. The intra-assay CV is 1.0% and the inter-assay CV is 1.3%.

      3. The detection range of the kit is 0.005 mmol/L.

      4. The average recovery rate of this kit is 102%.

       

      Notes

      1. The kit is for scientific research only.

      2. Instructions should be followed strictly, changes of operation may result in unreliable results.

      3. The validity of kit is 6 months.

      4. Do not use components from different batches of kit.


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