ELISA Kit for Defensin Beta 2 (DEFb2)

B-DF2; DEFB2; BD2; DEF-B2; HBD2; DEFB102; HBD-2; SAP1; DEFB4A; DEFB4; Skin-Antimicrobial Peptide 1

Specificity

This assay has high sensitivity and excellent specificity for detection of Defensin Beta 2 (DEFb2).
No significant cross-reactivity or interference between Defensin Beta 2 (DEFb2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Defensin Beta 2 (DEFb2) and the recovery rates were calculated by comparing the measured value to the expected amount of Defensin Beta 2 (DEFb2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 81-88 84
EDTA plasma(n=5) 86-94 89
heparin plasma(n=5) 82-98 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Defensin Beta 2 (DEFb2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Defensin Beta 2 (DEFb2) 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 Defensin Beta 2 (DEFb2) 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) 78-89% 84-92% 93-101% 80-96%
EDTA plasma(n=5) 83-97% 89-97% 80-102% 83-104%
heparin plasma(n=5) 79-91% 89-101% 80-91% 80-94%

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
Livestock Science The effect of Lactobacillus on the expression of porcine β-defensin-2 in the digestive tract of piglets ScienceDirect: S1871141311000126
Appl Environ Microbiol. Modulation of Porcine β-Defensins 1 and 2 upon Individual and Combined Fusarium Toxin Exposure in a Swine Jejunal Epithelial Cell Line Asm: Source
Plos Pathogens HIF-1α Is Essential for Effective PMN Bacterial Killing, Antimicrobial Peptide Production and Apoptosis in?Pseudomonas aeruginosa?Keratitis PubMed: PMC3715414
British Journal of Nutrition Identification of sociodemographic and clinical factors associated with the levels of human β-defensin-1 and human β-defensin-2 in the human milk of Han Chinese Pubmed: 24124699
University of Waikato The Assessment of Vitamin D, Antimicrobial Peptides and Procalcitonin in Bronchiectasis Waikato:Source
Vaccine On birth single dose live attenuated OPV and BCG vaccination induces gut cathelicidin LL37 responses at 6 week of age: A natural experiment Pubmed:25444792
International Journal of Dairy Technology Concentration characteristics of bovine β-defensin 1 and 2 in fresh bovine milk and infant formula Doi: 10.1111
Molecular Nutrition Food Research Green tea polyphenol epigallocatechin-3-gallate improves epithelial barrier function by inducing the productionof antimicrobial peptide pBD-1 and pBD-2 in monolayers of porcine intestinal epithelial IPEC-J2 cells. Pubmed:26991948
Respiratory Research Vitamin D and allergic airway disease shape the murine lung microbiome in a sex-specific manner. pubmed:27655266
Key Engineering Materials The Level of Inflammatory Cytokines and Antimicrobial Peptides after Composite Material Implantation and Contamination with Bacterial Culture. 16629795
Scientific Reports Measurements of AMPs in stratum corneum of atopic dermatitis and healthy skin–tape stripping technique Pubmed:29374283
Frontiers in Microbiology Swine-Derived Probiotic Lactobacillus plantarum Inhibits Growth and Adhesion of Enterotoxigenic Escherichia coli and Mediates Host Defense Pubmed:29997590
Journal of Dermatological Science Glyoxal-induced exacerbation of pruritus and dermatitis is associated with staphylococcus aureus colonization in the skin of a rat model of atopic dermatitis Pubmed:29496360
Catalog No. Related products for research use of Sus scrofa; Porcine (Pig) Organism species Applications (RESEARCH USE ONLY!)
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