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Measuring L-glutamine removing effect of L-glutamate acid!

L-Glutamine Assay Kit

Calprotectin consists of two proteins, S100A8 and S100A9, and is known as a biomarker of inflammatory bowel disease. Yamasa Corporation developed ELISA for measurement of rat S100A9 using an anti-S100A9 monoclonal antibody. Using this ELISA, we measured an S100A9 level in feces, collected from a rat model of dextran sulfate sodium (DSS) -induced ulcerative colitis (UC), and demonstrated an increased level of S100A9 in the early stage of inflammation, thereby providing an indicator of UC(1).
Using this ELISA, a rat S100A9 assay kit was produced.

Features of the kit

  • ELISA with a 96-well microtiter plate.
  • Rat S100A9 levels can be measured in a range of 3.75-240 ng/mL.
  • Measurement time:
    • primary reaction, 2 hours
    • secondary reaction, 1 hour
    • color reaction, 20 minutes
  • Rat serum and fecal*1 S100A9 levels can be measured.*2

*1: The measurement of fecal S100A9 levels requires extraction procedures. For details, see (1) of “Notes of measurement.”
*2: Samples need to be diluted before measurement. For details, see (2) of “Notes of measurement.”

Principles of measurement

Sandwich ELSIA with two kinds of mouse monoclonal anti-rat S100A9 antibody

Product contents

  • Product code: YMS-80126EX
  • Components:
  • Name Volume Quantity
    1 Antibody solid phase plate 96-well microtiter plate 1plate
    2 S100A9 standard solution 1 (3.75 ng/mL) 0.5ml 1vial
    3 S100A9 standard solution 2 ( 15 ng/mL) 0.5ml 1vial
    4 S100A9 standard solution 3 ( 60 ng/mL) 0.5ml 1vial
    5 S100A9 standard solution 4 (240 ng/mL) 0.5ml 1vial
    6 Diluent (3-fold concentrated solution) 25ml 1vial
    7 Washing stock (5-fold concentrated solution) 50ml 1vial
    8 Enzyme-labeled antibody 0.15ml 1vial
    9 Coloring solution A 11ml 1vial
    10 Coloring solution B 0.5ml 1vial
    11 Stop solution 11ml 1vial
  • Storage: 2-8°C
  • Shelf life: 18 months after the date of manufacture
  • Suggested retail price: ¥143,000

Measuring methods

  1. Dispense 100 µL of standard solution or sample onto an antibody solid phase plate.
     ↓ Allowed to stand for 2 hours at room temperature.
  2. After removal of the reaction solution, dispense 300 µL of wash solution into each well.
     ↓ Repeat the above procedures three times.
  3. After removal of the reaction solution, dispense 100 µL of labeled antibody into each well.
     ↓ Allowed to stand for 1 hour at room temperature.
  4. After removal of the reaction solution, dispense 300 µL of washing solution into each well.
     ↓ Repeat the above procedures three times.
  5. Dispense 100 µL of coloring solution into each well.
     ↓ Allowed to stand for 20 minutes at room temperature.
  6. Dispense 100 µL of stop solution into each well.
     ↓
  7. Measure absorbance of each well at 450 nm.

Notes of measurement

  1. Rat fecal samples were extracted as described below. Modify the procedures as needed.
    1. Preparation of a diluent for extraction
      A diluent for extraction was prepared by adding Triton X-100 to a solution, prepared by diluting the diluent in the kit with purified water, at a final concentration of 0.5% (w/v).
    2. Equipment
      Pestle, fecal collection tube (with a diameter that allows insertion of the pestle), etc.
    3. Extraction
      1. Weight the fecal collection tube in advance.
      2. Collect about 200 mg of feces into the tube.
      3. Weigh the tube, and subtracting the tube weight to determine the weight of the feces.
      4. Add 500 µL of diluent for extraction into a fecal collection tube. Allow the tube to stand for 15 minutes.
      5. Grind and stir the feces with a pestle.
      6. Collect all the residues, while washing the pestle with 500 µL of diluent for extraction.
      7. Centrifuge the residues at 12,000 g for five minutes at 4°C. Collect the supernatant into a new tube.
      8. Dilute the supernatant tenfold or above with a diluent for extraction.
      9. Determine S100A9 level in the supernatant using standard procedures to calculate the amount of S100A9 in 1 mg of feces.
  2. Before measurement, dilute the samples (rat sera and fecal extracts) about tenfold with the diluent in the kit. Dilute samples as needed if their concentrations are above the measurement ranges.
  3. For more information, contact Marketing & MR Section.

Product List

Product Name Cat# Quantity Price

L-Glutamine Assay Kit

YMS-80116 1KIT

¥ 80,000
$ 2134
€ 1600

References

Arima J et al., J Biochem 134, 805-812, 2003

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.