ELISA Kit for Trypsinogen Activation Peptide (TAP)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-103 99
EDTA plasma(n=5) 78-97 84
heparin plasma(n=5) 94-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Trypsinogen Activation Peptide (TAP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Trypsinogen Activation Peptide (TAP) 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 Trypsinogen Activation Peptide (TAP) 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) 92-105% 79-95% 82-97% 83-91%
EDTA plasma(n=5) 78-98% 90-103% 96-104% 93-104%
heparin plasma(n=5) 90-98% 79-105% 98-105% 92-105%

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 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.

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Magazine Citations
Diabetologia Diabetes aggravates acute pancreatitis and inhibits pancreas regeneration in mice SpringerLink: x4l01435t34452jr
Molecular Biology Reports Prior peritoneal lavage with hot 0.9% saline induces HSP70 expression and protects against cerulein-induced acute pancreatitis in rats Pubmed: 23096089
Journal of the Pancreas Case report of mannose-binding lectin (MBL) deficiency and postoperative sepsis and coagulopathy in a patient following total pancreatectomy for chronic pancreatitis. Pubmed:25262717
Journal of Gastroenterology A small molecule inhibitor of NFκB blocks ER stress and the NLRP3 inflammasome and prevents progression of pancreatitis pubmed:27418337
Indian Journal of Pharmaceutical Sciences Effects of Calcium Channel Blockers on Trypsinogen Activation and Severity of Cerulein-induced Acute Pancreatitis in Rats IJPS:Source
Tropical Journal of Pharmaceutical Research Investigation of non-pharmaceutical cures for acute pancreatitis induced by cerulein in rats
Pancreas The Regulatory Effect of the Kinase Inhibitor PD98059 on Autophagic Flux During Trypsinogen Activation in Pancreatic Acinar Cells Pubmed: 32011537
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