Instant ELISA Kit for Pepsin (PP)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 94-105 98
EDTA plasma(n=5) 84-98 93
heparin plasma(n=5) 93-101 96

Precision

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

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Instant 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) 85-99% 93-102% 87-96% 84-96%
EDTA plasma(n=5) 88-98% 78-101% 90-104% 85-99%
heparin plasma(n=5) 81-101% 78-93% 78-93% 84-91%

Stability

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 5 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 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 100µL standard or sample to each well. Incubate 30 minutes at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 30 minutes at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 10 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Radiology and Oncology LEVEL OF PEPSIN AND BILE ACIDS IN THE SALIVA OF PATIENTS WITH GLOTTIS T1 CARCINOMA Szd:Source
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
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