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Myeloperoxidase (MPO) Stool / Urine ELISA

Price:
$490.00
SKU:
MPO39-K01
Quantity:


product-insert-callout.png Myeloperoxidase (MPO) Stool / Urine ELISA

 

Myeloperoxidase (MPO) Stool / Urine ELISA Assay kit:
For Research Use Only
Size:  1x96 wells
Sensitivity:  0.3 ng/ml
Dynamic Range:  2 -512 ng/ml
Incubation Time:  2.5 hours
Sample Type:  Stool / Urine
Sample Size: 50 mg / 100ml
Controls Included

ELISA kit Developed and Manufactured in the USA

Intended Use
The Eagle Biosciences Myeloperoxidase (MPO) ELISA Assay kit is intended for use in the quantitative determination of human myeloperoxidase (MPO) levels in stool and urine samples. The Myeloperoxidase (MPO) ELISA Assay kit is useful for detecting elevated levels of myeloperoxidase in stool samples, which may serve as a sensitive predictor for inflammatory activities in the gastrointestinal tract.  The Eagle Biosciences MPO ELISA Assay kit is for research use only and not to be used in diagnostic procedures.
 
Assay Principle
This Myeloperoxidase (MPO) ELISA Assay kit is designed, developed and produced for the quantitative measurement of human myeloperoxidase in stool samples. The assay utilizes the two-site “sandwich” technique with selected antibodies that bind to different epitopes of myeloperoxidase.

Assay standards, controls and extracted patient samples are added directly to wells of a microtiter plate that is coated with antibody to myeloperoxidase. After an incubation period, the plate is washed and horseradish peroxidase (HRP) conjugated human myeloperoxidase antibody is added to each well. After the second incubation period, a “sandwich” of solid-phase monoclonal antibody - human myeloperoxidase – HRP conjugated antibody” is formed. The unbound antibodies and buffer matrix are removed in the subsequent washing step. For the detection of this immunocomplex, the well is then incubated with a substrate solution in a timed reaction and the absorbances are then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each microtiter well is directly proportional to the amount of human myeloperoxidase in the test sample. A standard curve is generated by plotting the absorbance versus the respective human myeloperoxidase concentration for each standard on a point-to-point or 4-parameter curve fitting. The concentration of human myeloperoxidase in test samples is determined directly from this standard curve.

  1. Add 100 µl of Standards, Controls and extracted patient samples into the designated microwells.
  2. Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 1.5 hr. ± 5 minutes at 400 to 450 rpm.
  3. Just prior to the end of the incubation time, dilute the proper amount of Tracer Antibody for the assay.
  4. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  5. Add 100 µL of above Tracer Antibody to each well.
  6. Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 45 minutes ± 5 minutes at 400 to 450 rpm.
  7. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  8. Add 100 µL of ELISA HRP Substrate into each of the wells.
  9. Cover the plate with aluminum foil to or other material to avoid exposure to light. Incubate plate static, at room temperature for 20 minutes.
  10. Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  11. Read the absorbance at 405 nm with reference filter at 620 nm or 650 nm.

Typical Standard Curve

fecal-mpo.png


REFERENCES

  • Saiki T.  Myeloperoxidase concentrations in the stool as a new parameter of inflammatory bowel disease. Kurume Med J. 1998;45:69-73.
  • Angriman I, et al. Enzymes in feces: useful markers of chronic inflammatory bowel disease. Clin Chim Acta. 2007;381(1):63-8.
  • Matheson NR, et al. Isolation and properties of human neutrophil myeloperoxidase. Biochemistry 1981;20:325-30.
  • Peterson CG, et al. Fecal levels of leukocyte markers reflect disease activity in patients with ulcerative colitis. Scand J Clin Lab Invest. 2007;67(8):810-20.
  • Weissman G, et al. Release of inflammatory mediators from stimulated neutrophils. N Engl J Med 1980;303:27-34.
  • Bos A et al. Characterization and quantification of the peroxidase in human monocytes. Biochem Biophys Acta 1978;525:37-44.