ELISA Kit for Defensin Beta 2 (DEFb2) Homo sapiens (Human) Sandwich ELISA

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

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  • ELISA Kit for Defensin Beta 2 (DEFb2) Packages (Simulation)
  • ELISA Kit for Defensin Beta 2 (DEFb2) Packages (Simulation)
  • ELISA Kit for Defensin Beta 2 (DEFb2) Results demonstration
  • SEA072Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 84-101 95
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 85-99 95

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) 79-91% 88-98% 82-96% 79-99%
EDTA plasma(n=5) 79-96% 78-102% 94-103% 80-91%
heparin plasma(n=5) 80-89% 79-102% 80-91% 79-90%

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.

ELISA Kit for Defensin Beta 2 (DEFb2)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Defensin Beta 2 (DEFb2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Defensin Beta 2 (DEFb2). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Defensin Beta 2 (DEFb2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Defensin Beta 2 (DEFb2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • The effect of Lactobacillus on the expression of porcine β-defensin-2 in the digestive tract of pigletsScienceDirect: S1871141311000126
  • Modulation of Porcine β-Defensins 1 and 2 upon Individual and Combined Fusarium Toxin Exposure in a Swine Jejunal Epithelial Cell LineAsm: Source
  • HIF-1α Is Essential for Effective PMN Bacterial Killing, Antimicrobial Peptide Production and Apoptosis in?Pseudomonas aeruginosa?KeratitisPubMed: PMC3715414
  • 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 ChinesePubmed: 24124699
  • The Assessment of Vitamin D, Antimicrobial Peptides and Procalcitonin in BronchiectasisWaikato:Source
  • On birth single dose live attenuated OPV and BCG vaccination induces gut cathelicidin LL37 responses at 6 week of age: A natural experimentPubmed:25444792
  • Concentration characteristics of bovine β-defensin 1 and 2 in fresh bovine milk and infant formulaDoi: 10.1111
  • 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
  • Vitamin D and allergic airway disease shape the murine lung microbiome in a sex-specific manner.pubmed:27655266
  • The Level of Inflammatory Cytokines and Antimicrobial Peptides after Composite Material Implantation and Contamination with Bacterial Culture.16629795
  • Measurements of AMPs in stratum corneum of atopic dermatitis and healthy skin–tape stripping techniquePubmed:29374283
  • Swine-Derived Probiotic Lactobacillus plantarum Inhibits Growth and Adhesion of Enterotoxigenic Escherichia coli and Mediates Host DefensePubmed:29997590
  • Glyoxal-induced exacerbation of pruritus and dermatitis is associated with staphylococcus aureus colonization in the skin of a rat model of atopic dermatitisPubmed:29496360
  • Caprylic acid and nonanoic acid upregulate endogenous host defense peptides to enhance intestinal epithelial immunological barrier function via histone deacetylase …Pubmed: 30342347
  • Influence of Antibiotic-Impregnated Biomaterials on Inflammatory Cytokines
  • Bovine fetal mesenchymal stem cells exert antiproliferative effect against mastitis causing pathogen Staphylococcus aureusPubmed: 30975214
  • Damage-Associated Molecular Patterns and Myeloid-Derived Suppressor Cells in Bronchoalveolar Lavage Fluid in Chronic Obstructive Pulmonary Disease Patients
  • Toll-like receptor 5-mediated IL-17C expression in intestinal epithelial cells enhances epithelial host defense against F4+ ETEC infectionPubmed: 31221216
  • Fat Encapsulation Reduces Diarrhea in Piglets Partially by Repairing the Intestinal Barrier and Improving Fatty Acid Transport33375218
  • Human beta defensin levels and vaginal microbiome composition in post-menopausal women diagnosed with lichen sclerosus34362937
  • Regulatory T cells, damage-associated molecular patterns, and myeloid-derived suppressor cells in bronchoalveolar lavage fluid interlinked with chronic obstructive …Pubmed:35687771

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