Multiplex Assay Kit for Trypsinogen Activation Peptide (TAP) ,etc. by FLIA (Flow Luminescence Immunoassay)

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-101 95
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 81-94 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Trypsinogen Activation Peptide (TAP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 88-102% 78-98% 83-97% 90-104%
EDTA plasma(n=5) 92-104% 89-98% 96-104% 78-101%
heparin plasma(n=5) 85-104% 78-96% 79-104% 91-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:TAP) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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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