ELISA Kit for Defensin Beta 1 (DEFb1)

B-DF1; DEFB1; BD1; DEF-B1; DEFB101; HBD1

Specificity

This assay has high sensitivity and excellent specificity for detection of Defensin Beta 1 (DEFb1).
No significant cross-reactivity or interference between Defensin Beta 1 (DEFb1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Defensin Beta 1 (DEFb1) and the recovery rates were calculated by comparing the measured value to the expected amount of Defensin Beta 1 (DEFb1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-98 94
EDTA plasma(n=5) 79-102 91
heparin plasma(n=5) 81-99 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Defensin Beta 1 (DEFb1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Defensin Beta 1 (DEFb1) 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 1 (DEFb1) 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) 80-98% 90-98% 87-99% 79-101%
EDTA plasma(n=5) 90-101% 93-101% 82-93% 90-98%
heparin plasma(n=5) 97-105% 79-95% 82-101% 78-89%

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
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
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
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
Frontiers in Microbiology Swine-Derived Probiotic Lactobacillus plantarum Inhibits Growth and Adhesion of Enterotoxigenic Escherichia coli and Mediates Host Defense Pubmed:29997590
International Immunopharmacology Caprylic acid and nonanoic acid upregulate endogenous host defense peptides to enhance intestinal epithelial immunological barrier function via histone deacetylase … Pubmed: 30342347
cyberleninka EFFECT OF NIOSOMAL ANTIMICROBIAL PEPTIDE HBD-1 ON THE HEALING RATE OF INFECTED WOUNDS IN RATS
Fundamental & clinical pharmacology Effect of antimicrobial peptides HNP‐1 and hBD‐1 on Staphylococcus aureus strains in vitro and in vivo Pubmed: 31313350
Cytokine Levels of pro-and anti-inflammatory cytokines in cystic fibrosis patients with or without gingivitis Pubmed: 31927460
Animals Fat Encapsulation Reduces Diarrhea in Piglets Partially by Repairing the Intestinal Barrier and Improving Fatty Acid Transport 33375218
Sci Rep Human beta defensin levels and vaginal microbiome composition in post-menopausal women diagnosed with lichen sclerosus 34362937
J Mol Histol Isolation, culture, and identification of ceruminous gland cells Pubmed:35113280
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