Multiplex Assay Kit for Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array)
B-DF2; DEFB2; BD2; DEF-B2; HBD2; DEFB102; HBD-2; SAP1; DEFB4A; DEFB4; Skin-Antimicrobial Peptide 1
Overview
Properties
- Product No.CMA072Mu
- Organism SpeciesMus musculus (Mouse) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryInfection immunity
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Recovery
Matrices listed below were spiked with certain level of recombinant Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 78-104 | 87 |
| EDTA plasma(n=5) | 82-104 | 85 |
| heparin plasma(n=5) | 82-97 | 91 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Defensin Beta 2 (DEFb2) ,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 Defensin Beta 2 (DEFb2) ,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 Defensin Beta 2 (DEFb2) ,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) | 82-96% | 80-104% | 80-92% | 90-103% |
| EDTA plasma(n=5) | 95-103% | 86-101% | 82-99% | 91-101% |
| heparin plasma(n=5) | 78-101% | 91-101% | 78-94% | 88-98% |
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.
Test principle
Anti-Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Defensin Beta 2 (DEFb2) ,etc. by CBA (Cytometric Bead Array) in the sample and corresponding value of MFI (Median Fluorescence Intensity). Flow Cytometer was used to determine and calculate the sample concentration.
Giveaways
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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
