PGC ELISA Kit

Catalog No. ABIN1305146

Product Details PGC ELISA Kit

Target: PGC (Pepsinogen C (PGC))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.5-100 ng/mL

Minimum Detection Limit: 0.5 ng/mL

Application: ELISA

Purpose: This ELISA (enzyme-linked immunosorbent assay) kit is intended for the quantitative determination of human pepsinogen II levels in serum. Determination of human serum pepsinogen II level would allow calculation of the ratio of pepsinogen I/II, which was reported to be a useful tool in the aid of diagnosing the functional states of acid-secreting gastric mucosa.

Brand: ED™

Sample Type: Serum

Analytical Method: Quantitative

Specificity: This assay measures human pesinogen II without any cross-reaction to human pesinogen I.

Components: 1. Streptavidin Coated Microplate
Two vials each contain lyophilized human pepsinogen II in a bovine serum albumin-based matrix with a non-azide preservative. Refer to vials for exact concentration range for each control.

Both controls should be reconstituted with DI-water and stored at -20 °C or below after the first use with up to 3 freeze cycles.

Material not included: 1. Precision single channel pipettes capable of delivering 20 µL, 25 µL, 100 µL, and 1000 µL, etc.
2. Repeating dispenser suitable for delivering 100 µL.
3. Disposable pipette tips suitable for above volume dispensing.
4. Disposable 12 x 75 mm or 13 x 100 glass tubes
5. Disposable plastic 1000 mL bottle with caps.
6. Aluminum foil.
7. Deionized or distilled water.
8. Plastic microtiter well cover or polyethylene film.
9. ELISA multichannel wash bottle or automatic (semi-automatic) washing system.
10. Spectrophotometric microplate reader capable of reading absorbance at 450 nm

Application Details

Sample Volume: 50 μL

Assay Time: 4 h

Plate: Pre-coated

Protocol: Assay standards, controls and patient serum samples containing human pepsinogen II are added directly to microtiter wells of microplate that was coated with streptavidin. Simultaneously, a biotinylated antibody and a horseradish peroxidase-conjugated antibody are added to each well. After the first incubation period, the wall of microtiter well captures the biotinylated antibody as well as an immunocomplex 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.

Reagent Preparation:

(1) Prior to use allow all reagents to come to room temperature. Reagents from different kit lot numbers should not be combined or interchanged.
(2) ELISA Wash Concentrate must be diluted to working solution prior use. Please see REAGENTS section for details.
(3) Reconstitute all assay standards and controls by adding 0.5 mL of deminerialized water to the vial of standard level 1 and 0.5 mL deminerialized water to the vials of standard levels 2 - 6 and controls 1 & 2. Allow the standards and controls to sit undisturbed for 10 minutes, and then mix well by gentle vortexing. Make sure that all solid is dissolved completely prior to use. These reconstituted standards and controls must be stored at - 10 C or below. Do not exceed 3 freeze-thaw cycles.
(4) Place a sufficient number of Streptavidin coated microwell strips in a holder to run human pepsinogen II standards, controls and unknown samples in duplicate.
(5) Test Configuration
(6) Prepare working Tracer Antibody and Capture Antibody mixture by 1:21 fold dilution of the Pepsinogen II Tracer Antibody and the Pepsinogen II Capture Antibody with the Tracer Antibody Diluent . For each strip, it is required to mix 1 mL of Tracer Antibody Diluent with the addition of 50 L of Tracer Antibody and 50 L Capture Antibody in a clean test tube or vial. Following is a table that outlines the relationship of strips used and antibody mix prepared.

Sample Collection: Only 100 µL of human serum is required for human pepsinogen II measurement in duplicate. No special preparation of individual is necessary prior to specimen collection. However, a 10 hour fasting serum sample is recommended for the test. Whole blood should be collected and must be allowed to clot for minimum 30 minutes at room temperature before the serum is separated by centrifugation (850 ? 1500xg for 10 minutes). The serum should be separated from the clot within three hours of blood collection and transferred to a clean test tube. Serum samples should be stored at -20 °C or below until measurement. Avoid more than three times freeze-thaw cycles of specimen.

Assay Procedure:

(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) Incubate plate with initial shaking for 1 minutes and further incubation at 37 °C for 60 minutes.
(4) Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents.
(5) Add 100 µL of ELISA HRP Substrate into each of the wells
(6) Incubate plate at 37 °C for 18 minutes.
(7) Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
(8) Read the absorbance at 450 nm.

Calculation of Results:

1. Calculate the average absorbance for each pair of duplicate test results.
   2. Subtract the average absorbance of the STD 1 (0 ng/mL) from the average absorbance of all other readings to obtain corrected absorbance.
   3. The standard curve is generated by the corrected absorbance of all standard levels on the ordinate against the standard concentration on the abscissa using point-to-point or log-log paper. Appropriate computer assisted data reduction programs may also be used for the calculation of results.
   4. It is recommended to use following curve fits: (1) Point-to-Point, or (2) 4-Parameter or (3) CubicSpline. The human pepsinogen II concentrations for the controls and patient samples are read directly from the standard curve using their respective corrected absorbance.

Assay Precision: The intra-assay precision is validated by measuring two samples in a single assay with 16 replicate determinations. The inter-assay precision is validated by measuring two samples in duplicate in 12 individual assays.

Restrictions: For Research Use only

Handling

Precaution of Use: The reagents must be used in research laboratory and is for research use only. Source material for reagents containing bovine serum albumin was derived in the contiguous 48 United States. It was obtained only from healthy donor animals maintained under veterinary supervision and found free of contagious diseases. Wear gloves while performing this assay and handle these reagents as if they are potentially infectious. Avoid contact with reagents containing TMB, hydrogen peroxide, or sulfuric acid. TMB may cause irritation to skin and mucous membranes and cause an allergic skin reaction. TMB is a suspected carcinogen. Sulfuric acid may cause severe irritation on contact with skin. Do not get in eyes, on skin, or on clothing. Do not ingest or inhale fumes. On contact, flush with copious amounts of water for at least 15 minutes. Use Good Laboratory Practices.

Storage: 4 °C

Target Details for PGC

Target: PGC (Pepsinogen C (PGC))

Alternative Name: Pepsinogen II (PGC Products)

Synonyms: PEPC ELISA Kit, PGII ELISA Kit, 2210410L06Rik ELISA Kit, Upg-1 ELISA Kit, Upg1 ELISA Kit, CPGC ELISA Kit, gastricsin ELISA Kit, PG1 ELISA Kit, Pg-1 ELISA Kit, PGNC ELISA Kit, PGC ELISA Kit, Gastricsin ELISA Kit, progastricsin (pepsinogen C) ELISA Kit, progastricsin ELISA Kit, PGC ELISA Kit, Pgc ELISA Kit

Target Type: Antibody

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 and mucous neck cells, but also by clear mucous cells of antrum, etc. The clinical applications of measuring pepsinogen I and pepsinogen II are a 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 serum pepsinogen I levels falling 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 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 use 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.

Molecular Weight: 42 kDa