Multiplex Assay Kit for Polymeric Immunoglobulin Receptor (PIGR) ,etc. by FLIA (Flow Luminescence Immunoassay)

SC; Secretory Component; Hepatocellular carcinoma-associated protein TB6

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-96 92
EDTA plasma(n=5) 95-103 98
heparin plasma(n=5) 79-104 90
sodium citrate plasma(n=5) 81-89 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Polymeric Immunoglobulin Receptor (PIGR) ,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 Polymeric Immunoglobulin Receptor (PIGR) ,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 Polymeric Immunoglobulin Receptor (PIGR) ,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) 94-101% 96-104% 84-97% 82-99%
EDTA plasma(n=5) 87-103% 81-102% 86-93% 96-105%
heparin plasma(n=5) 79-97% 79-101% 99-105% 95-102%
sodium citrate plasma(n=5) 96-104% 85-92% 85-105% 98-105%

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#:PIGR) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Journal of Proteome Research Sputum proteomics identifies elevated PIGR levels in smokers and mild to moderate COPD ACS: pr2006395
Molecular & Cellular Proteomics Integrated Proteomic Profiling of Cell Line Conditioned Media and Pancreatic Juice for the Identification of Pancreatic Cancer Biomarkers PubMed: PMC3205865
BMC Complementary and Alternative Medicine Identification of plasma protein markers common to patients with malignant tumour and Abnormal Savda in Uighur medicine: a prospective clinical study Pubmed:25652121
British Journal of Cancer Identification of a novel serum biomarker for pancreatic cancer, C4b-binding protein α-chain (C4BPA) by quantitative proteomic analysis using tandem mass tags. pubmed:27657339
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