Citrid Acid (CA) Colorimetric Assay Kit

    • Biochemical Assay Kit-Elabscience
    <
    >
    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K351

      Size:
      • 50 Assays
      Qty:
      - +
      Price: $220

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry (Visible range)

      Valid period: 3 months

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

      Add to cart Compare Bulk request Manual


      Application

      This kit can be used to measure citric acid (CA) content in serum, plasma, tissue, mitochondria and other samples. This kit (50 Assays) can detect 48 samples.

       

      Detection significance

      CA is a common organic acid in organisms and an important food flavor substance. In addition, CA is the product of the first step of the tricarboxylic acid cycle.

       

      Detection principle

      Under acidic conditions, citric acid reduced Cr6+ to form Cr3+, which had characteristic absorption peak at 545 nm. And the citric acid content in the sample can be calculated by measuring the increasing of absorbance value at 545 nm.



      Experimental instrument

      Test tube, Mortar, High-precision transferpettor, Vortex mixer, Refrigerated Centrifuge, Spectrophotometer (240 nm)

       

      Extraction of citric acid:

      1. Serum / plasma samples:

      Take 0.1 mL sample and add 0.9 mL reagent 1, mix fully, then centrifuge at 11000 g for 10 min at 4. Take the supernatant and preserve it on ice for detection.

      2. Preparation of tissue samples:

      Add reagent 1 into the tissue sample according to the ratio of Weight (g): reagent 1 (mL) =1:5~10 (it is recommended to add 1 mL of reagent 1 into 0.1 g of tissue), then homogenize the sample on ice. Centrifuge the tissue homogenate at 11000 g for 10 min at 4. Take the supernatant and preserve it on ice for detection.

      3. Preparation of mitochondria:

      Add reagent 1 into the tissue sample according to the ratio of Weight (g): reagent 1 (mL) =1:5~10 (it is recommended to add 1 mL of reagent 1 into 0.1 g of tissue), then homogenize the sample on ice. Centrifuge the tissue homogenate at 600 g for 5 min at 4. Take the supernatant to another EP tube, centrifuge the supernatant at 11000 g for 10 min at 4. Discard the supernatant (the supernatant can be used for the determination of CA content in cytoplasm.). Add 200 μL reagent 2 and 2 μL reagent 3 to the sediment and dissolve fully. Centrifuge the solution at 11000 g for 10 min at 4. Take the supernatant and preserve it on ice for detection.

      4. Preparation of bacteria or fungi samples:

      Bacteria or fungi: Collect the bacteria or fungi samples into a centrifuge tube, then centrifuge the sample and discard the supernatant. Add Reagent 1 into the sediment according to the ratio of Bacteria or Cells number (104): Reagent 1 (mL) =500~1000: 1 (it is recommended to add 1 mL of Reagent 1 into 5×106 bacteria or fungi), then treat the sample with sonication on ice (power: 300W, 3 seconds/time, interval for 7 seconds, total time 3 min). Centrifuge at 11000 g for 10 min at 4. Take the supernatant and preserve it on ice for detection.

       

      Detection procedures

      1. Preheat the Spectrophotometer or Microplate Reader for 30 min, then adjust the wavelength at 545 nm and set to zero with distilled water.

      2. Incubate the reagent 1 at 30 for 30 min before detection.

      3. Operation table

       

      Blank tube

      Standard tube

      Sample tube

      Double distilled water(μL)

      100

       

       

      Standard solution (μL)

       

      100

       

      Sample (μL)

       

       

      100

      Reagent 1 (μL)

      700

      700

      700

      Reagent 4 (μL)

      100

      100

      100

      Reagent 5 (μL)

      100

      100

      100

      Mix fully and stand at room temperature for 30 min. Set the spectrophotometer to zero with double distilled water and measure the OD value at 545 nm and recorded as A.

       

      Note:

      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.

      5. All processes of sample preparation need to be carried out on ice

      6. Reagent 5 is a carcinogenic substance. During the experiment, gloves should be worn to avoid splash of reagents on the skin.

      7. Citric acid extraction solution cannot be used for the determination of protein content. If the content of protein needs to be determined, tissues should be taken separately (E-BC-K318, E-BC-K168, E-BC-K165).



      • Show all (0)
      • Reviews (0)
      • Q&A (0)
      ... Show All Show Less
      MSDS for ELISA

      Browsing History


        People Also Bought