Multiplex Assay Kit for Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array)

REG-III; PAP1B; PAPIB; UNQ429; Pancreatitis-associated protein 1B; Regenerating islet-derived protein III-gamma

Specificity

This assay has high sensitivity and excellent specificity for detection of Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-89 85
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 79-102 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) 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 Regenerating Islet Derived Protein 3 Gamma (REG3g) ,etc. by CBA (Cytometric Bead Array) 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) 91-102% 84-97% 98-105% 95-104%
EDTA plasma(n=5) 87-96% 99-105% 99-105% 84-97%
heparin plasma(n=5) 82-98% 84-103% 98-105% 84-101%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
16 Innate Stat3-mediated induction of the antimicrobial protein Reg3γ is required for host defense against MRSA pneumonia PubMed: PMC3600913
Biology of Blood and Marrow Transplatation Reciprocal Expression of Enteric Antimicrobial Proteins in Intestinal Graft-Versus-Host Disease Pubmed: 23927965
Nature Comunication Hyperglycaemia inhibits REG3A expression to exacerbate TLR3-mediated skin inflammation in diabetes. pubmed:27830702
Gastroenterology Enteric Delivery of Regenerating Family Member 3 alpha Alters the Intestinal Microbiota and Controls Inflammation in Mice With Colitis Pubmed:29133078
Kidney International Interleukin-6/Stat3 signaling has an essential role in the host antimicrobial response to urinary tract infection Pubmed:29475562
Journal of Immunology Research Role of TLR5 in the Translocation and Dissemination of Commensal Bacteria in the Intestine after Traumatic Hemorrhagic Shock 34840990
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