Pepsinogen II ELISA Kit

$480.00

The Eagle Biosciences Human Pepsinogen II ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human pepsinogen II levels in serum. The Eagle Biosciences Human Pepsinogen II ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: PP231-K01 Categories: , ,

Pepsinogen II ELISA Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.1 ng/mL
Dynamic Range: 6.3 – 100 ng/mL
Incubation Time: 2.5 hours
Sample Type: Serum
Sample Size: 50 µL

Controls Included

Product Developed and Manufactured in the USA

Additional Information

Assay Background

Pepsinogen consists of a single polypeptide chain of 375 amino acids with an average molecular weight of 42 kDa. Pepsinogen I is synthesized at gastric chief cells and mucous neck cells, while pepsinogen II is produced not only by gastric chief cells, mucous neck cells, but also by clear mucous cells of antrum and Brunner’s glands in the proximal duodenum, etc. The clinical applications of measuring pepsinogen I and pepsinogen II are of useful aid in diagnosing severe atrophic gastritis and stomach cancer. It was suggested that the measurement of serum pepsinogens served as a “serological biopsy” for predicting the presence of atrophic gastritis or superficial gastritis.

Atrophic Gastritis: It was found that a serum pepsinogen I level failed to less than 20 ng/ml was highly specific for severe atrophic gastritis. It is also observed that serum pepsinogen I levels fell with increasing severity of mucosal damage in atrophic gastritis. The diagnostic sensitivity and specificity of serum pepsinogen I level for advanced atrophic corpus gastritis are about 92% and 90% respectively. On the other hand, the decrease in serum pepsinogen I levels in patients with pernicious anemia and atrophic gastritis was found to be associated with normal or raised pepsinogen II levels. Therefore, a pepsinogen I/pepsinogen II ratio is significantly lower than those with superficial gastritis or normal remnant mucosa.

Stomach Cancer: Low serum pepsinogen I levels as well as low ratio of pepsinogen I/II were found in patients with gastric cancer, with a threefold higher incidence. Other studies have concluded that low serum pepsinogen I levels may identify persons at increased risk for intestinal types of stomach cancer.

Duodenal Ulcer: A low serum pepsinogen I level can exclude a diagnosis of duodenal ulcer. Although a high pepsinogen I level has less clinical useful for establishing the diagnosis of a duodenal ulcer, the combination of hypergastrinemia and a highly elevated serum pepsinogen I strongly suggests the possibility of the Zollinger-Ellison syndrome.

Assay Principle

The Eagle Biosciences Human Pepsinogen II ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human pepsinogen II level in serum sample. The assay utilizes the two-site “sandwich” technique with two selected monoclonal antibodies that bind to different epitopes of human pepsinogen II without any cross-reaction to human pepsinogen I.

Assay standards, controls and patient serum samples containing human pepsinogen II is added directly to microtiter wells of microplate that was coated with a streptavidin. Simultaneously, a biotinylated antibody and a horseradish peroxidase conjugated antibody is added to each well. After the first incubation period, on the wall of microtiter well captures the biotinylated antibody as well as an immuno complex in the form of “streptavidin – biotin-antibody – pepsinogen II – HRP-antibody”. Unbound proteins as well as unbound HRP conjugated antibody in each microtiter well are removed in the subsequent washing step. The well is incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the tracer antibody bound to the pepsinogen II on the wall of the microtiter well is directly proportional to the amount of pepsinogen II in the sample. A standard curve is generated by plotting the absorbance versus the respective human pepsinogen II concentration for each standard on Point-to-Point, CubicSpline or 4-Parameter plot. The concentration of human pepsinogen II in test samples is determined directly from this standard curve.

  1. Add 50 µL of standards, controls and patient serum samples into the designated microwell.
  2. Add 100 µL of above antibody mixture to each well
  3. Mix gently and cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
  4. Incubate plate at room temperature for 2 hours.
  5. Remove the aluminum foil and plate sealer. Aspirate the contents of each well. 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.
  6. Add 100 µL of ELISA HRP Substrate into each of the wells.
  7. Cover the plate with one new plate sealer and also with aluminum foil to avoid exposure to light.
  8. Incubate plate at room temperature for 20 minutes (This incubation period may be reduced to 8 – 15 min if a lower OD reading is demanded to fit to the plate readers specification)
  9. Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  10. Read the absorbance at 450 nm within 10 minutes in a microplate reader.

Typical Assay Curve

Pepsinogen II ELISA Kit Standard Curve

Manual

Product Manual