ELISA Kit for Pepsin (PP)


This assay has high sensitivity and excellent specificity for detection of Pepsin (PP).
No significant cross-reactivity or interference between Pepsin (PP) and analogues was observed.


Matrices listed below were spiked with certain level of recombinant Pepsin (PP) and the recovery rates were calculated by comparing the measured value to the expected amount of Pepsin (PP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-102 97
EDTA plasma(n=5) 81-99 91
heparin plasma(n=5) 88-99 94


Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pepsin (PP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pepsin (PP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Pepsin (PP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-99% 92-105% 86-94% 78-101%
EDTA plasma(n=5) 83-97% 82-101% 86-99% 95-102%
heparin plasma(n=5) 97-105% 85-94% 97-104% 97-104%


The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.



Magazine Citations
Radiology and Oncology Higher levels of total pepsin and bile acids in the saliva as a possible risk factor for early laryngeal cancer Pubmed:Pmc4362607
Clin Otolaryngol Pepsin and bile acids in saliva in patients with laryngopharyngeal reflux – a prospective comparative study PubMed: 25516364
journal of pediatric gastroenterology and nutrition Does Positioning Affect Tracheal Aspiration of Gastric Content in Ventilated Infants? PubMed: 25313850
Journal of Voice Detecting Laryngopharyngeal Reflux by Immunohistochemistry of Pepsin in the Biopsies of Vocal Fold Leukoplakia pubmed:28756936
Sensors (Basel) Label-Free Detection of Salivary Pepsin Using Gold Nanoparticle/Polypyrrole Nanocoral Modified Screen-Printed Electrode Pubmed:29882917
The Tohoku Journal of Experimental Medicine Association between Pepsin in Bronchoalveolar Lavage Fluid and Prognosis of Chronic Fibrosing Interstitial Lung Disease
Respiratory Research Pulmonary aspiration in preschool children with cystic fibrosis Pubmed: 30558606
Tanaffos Association of Bile Acid and Pepsin Micro-aspiration with Chronic Obstructive Pulmonary Disease Exacerbation Pubmed: 31423141
Sensors Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux Pubmed: 31935973
BIOSCIENCE REPORTS Presence of pepsin in laryngeal tissue and saliva in benign and malignant neoplasms Pubmed: 33103719
OTOLARYNGOLOGY-HEAD AND NECK SURGERY Relationship Between Pepsin Expression and Dysplasia Grade in Patients With Vocal Cord Leukoplakia Pubmed: 32692278
American Journal of Otolaryngology Association of pepsin and DNA damage in laryngopharyngeal reflux-related vocal fold polyps Pubmed: 32889371
The relationship between laryngopharyngeal reflux based on pepsin value and clinical characteristics of laryngeal cancer patients
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