Wide-range ELISA Kit for Regenerating Islet Derived Protein 3 Gamma (REG3g)

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 Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g).
No significant cross-reactivity or interference between Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) and the recovery rates were calculated by comparing the measured value to the expected amount of Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-102 91
EDTA plasma(n=5) 94-104 98
heparin plasma(n=5) 82-96 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) 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 Wide-range Regenerating Islet Derived Protein 3 Gamma (REG3g) 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) 80-93% 93-101% 94-102% 80-90%
EDTA plasma(n=5) 93-101% 87-94% 81-97% 80-97%
heparin plasma(n=5) 94-105% 78-101% 89-96% 79-99%

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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 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
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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