Sialic Acid (SA) Colorimetric Assay Kit

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

      Catalog number:E-BC-K068

      Size:
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $150

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry (Visible range)

      Valid period: 3 months

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

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      Application

      This kit can be used to measure the Sialic Acid (SA) content in serum, plasma, tissue, saliva and other samples. 

       

      Detection significance

      Sialic acid is a general name of derivatives with nine carbon glycosylneuraminic acid. Sialic acid is widely distributed in animal tissues, mainly in glycoprotein and ganglioside. The main function of sialic acid is to participate in various recognition processes between cells and molecules. N-acetyl neuraminic acid, an important molecule in biometrics, is the representative of sialic acid family.

       

      Detection principle

      Sialic acid forms a purplish red complex with methyl resorcinol in the presence of oxidant. And the absorbance conforms to Lambert-Beer's law. The content of sialic acid can be calculated by measuring the OD value at 560 nm.

      When detect tissue and cell samples, the protein concentration of the sample needs to be detected in addition (E-BC-K165, E-BC-K168, E-BC-K318 are recommended).

      Experimental instrument

      Tube, Micropipette, Vortex mixer, Centrifuge, Spectrometer (560 nm), Water bath

       

      The preparation of sample

      1.         Serum (plasma) sample: detect directly.

      2.         Saliva sample: saliva was collected 30 min after gargling with clear water and centrifuged at 10000 g at 4 for 5 min. Then take the supernatant for detection.

      3.         10% tissue homogenate: Weigh the tissue, add PBS (0.01 M, pH 7.4) in a 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 for detection.

      Operation steps

      It is recommended to take 2~3 samples which expected large difference to do pre-experiment before formal experiment.

      1. Blank tube: For serum (plasma) and saliva samples, add 0.1 mL double distilled water into a 5 mL EP tube. For tissue sample, add 0.2 mL double distilled water into a 5 mL EP tube

      Standard tube: For serum (plasma) and saliva samples, add 0.1 mL of 1 mmol/L SA standard into a 5 mL EP tube. For tissue sample, add 0.1 mL of 1 mmol/L SA standard and 0.1 mL of double distilled water into a 5 mL EP tube.

      Sample tube: For serum (plasma) and saliva samples, add 0.1 mL sample into a 5 mL EP tube. For tissue sample, add 0.2 mL sample into a 5 mL EP tube.

      2. Add 4 mL of Reagent 2 into each tube.

      3. Mix fully with a vortex mixer, then tight the tube with plastic membrane (make a small hole on the membrane). Incubate the tubes at 100 for 15 min.

      4. Take out the tubes and cool with running water. Centrifuge at 2325 g for 10 min.

      5. Set spectrophotometry to zero with ddH2O Take the supernatant and measure the OD values of each tube at 560 nm with 1 cm diameter cuvette.

       

      Note: It can be refer to the following operating table.

      (1) For serum (plasma) and saliva sample:

       

      Blank tube

      Standard tube

      Sample tube

      Double-distilled water (mL)

      0.1

       

       

      1 mmol/L SA standard (mL)

       

      0.1

       

      Sample (mL)

       

       

      0.1

      Reagent 2 (mL)

      4.0

      4.0

      4.0

      Mix fully with a vortex mixer, then tight the tube with plastic membrane (make a small hole on the membrane). Incubate the tubes at 100 for 15 min. Take out the tubes and cool with running water. Centrifuge at 2325 g for 10 min. Set spectrophotometry to zero with ddH2O Take the supernatant and measure the OD values of each tube at 560 nm with 1 cm diameter cuvette.

      (2) For tissue sample:

       

      Blank tube

      Standard tube

      Sample tube

      Double-distilled water (mL)

      0.2

      0.1

       

      1 mmol/L SA standard (mL)

       

      0.1

       

      Sample (mL)

       

       

      0.2

      Reagent 2 (mL)

      4.0

      4.0

      4.0

      Mix fully with a vortex mixer, then tight the tube with plastic membrane (make a small hole on the membrane). Incubate the tubes at 100 for 15 min. Take out the tubes and cool with running water. Centrifuge at 2325 g for 10 min. Set spectrophotometry to zero with ddH2O Take the supernatant and measure the OD values of each tube at 560 nm with 1 cm diameter cuvette.


      Technical parameters

      1. The sensitivity of the kit is 0.022 mmol/L.

      2. The intra-assay CV is 3.9 % and the inter-assay CV is 7.1%.

      3. The recovery of the kit is 95%.

      4. The linear range of the kit is 0.022-7 mmol/L.

       

      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 3 months.

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


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