防御素β2(DEFb2)檢測試劑盒(酶聯免疫吸附試驗法)

ELISA Kit for Defensin Beta 2 (DEFb2)

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

  • 防御素β2(DEFb2)檢測試劑盒(酶聯免疫吸附試驗法) 產品包裝(模擬)
  • 防御素β2(DEFb2)檢測試劑盒(酶聯免疫吸附試驗法) 產品包裝(模擬)
  • 防御素β2(DEFb2)檢測試劑盒(酶聯免疫吸附試驗法) 實驗結果圖
  • SEA072Po.jpg 標準曲線圖
  • Certificate 通過ISO 9001、ISO 13485質量體系認證

特異性

本試劑盒用于檢測防御素β2(DEFb2),經檢測與其它相似物質無明顯交叉反應。
由于受到技術及樣本來源的限制,不可能完成對所有相關或相似物質交叉反應檢測,因此本試劑盒有可能與未經檢測的其它物質有交叉反應。

回收率

分別于定值血清及血漿樣本中加入一定量的防御素β2(DEFb2)(加標樣品),重復測定并計算其均值,回收率為測定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 81-88 84
EDTA plasma(n=5) 86-94 89
heparin plasma(n=5) 82-98 93

精密度

精密度用樣品測定值的變異系數CV表示。CV(%) = SD/mean×100
批內差:取同批次試劑盒對低、中、高值定值樣本進行定量檢測,每份樣本連續測定20 次,分別計算不同濃度樣本的平均值及SD值。
批間差:選取3個不同批次的試劑盒分別對低、中、高值定值樣本進行定量測定,每個樣本使用同一試劑盒重復測定8次,分別計算不同濃度樣本的平均值及SD值。
批內差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內加入適量的防御素β2(DEFb2),并倍比稀釋成1:2,1:4,1:8,1:16的待測樣本,線性范圍即為稀釋后樣本中防御素β2(DEFb2)含量的測定值與理論值的比率。

樣本 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%

穩定性

經測定,試劑盒在有效期內按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對試劑盒破壞前后檢測值的影響,實驗室的環境條件需盡量保持一致,尤其是實驗室內溫度、濕度及溫育條件。其次由同一實驗員來進行操作可減少人為誤差。

實驗流程

1. 實驗前標準品、試劑及樣本的準備;
2. 加樣(標準品及樣本)100µL,37°C孵育1小時;
3. 吸棄,加檢測溶液A100µL,37°C孵育1小時;
4. 洗板3次;
5. 加檢測溶液B100µL,37°C孵育30分鐘;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分鐘;
8. 加終止液50µL,立即450nm讀數。

實驗原理

將防御素β2(DEFb2)抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標準品或標本,其中的防御素β2(DEFb2)與連接于固相載體上的抗體結合,然后加入生物素化的防御素β2(DEFb2)抗體,將未結合的生物素化抗體洗凈后,加入HRP標記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過氧化物酶的催化下轉化成藍色,并在酸的作用下轉化成最終的黃色。顏色的深淺和樣品中的防御素β2(DEFb2)呈正相關。用酶標儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。

相關產品

編號 適用物種:Sus scrofa; Porcine (Pig,豬) 應用(僅供研究使用,不用于臨床診斷!)
RPA072Po01 防御素β2(DEFb2)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAA072Po01 防御素β2(DEFb2)多克隆抗體 WB; IHC; ICC; IP.
MAA072Po21 防御素β2(DEFb2)單克隆抗體 WB; IHC; ICC; IP.
SEA072Po 防御素β2(DEFb2)檢測試劑盒(酶聯免疫吸附試驗法) Enzyme-linked immunosorbent assay for Antigen Detection.

參考文獻

雜志 參考文獻
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]
International Immunopharmacology Caprylic acid and nonanoic acid upregulate endogenous host defense peptides to enhance intestinal epithelial immunological barrier function via histone deacetylase?… [Pubmed: 30342347]
拉脫維亞科學院院刊 Influence of Antibiotic-Impregnated Biomaterials on Inflammatory Cytokines []
Veterinary?Research Bovine fetal mesenchymal stem cells exert antiproliferative effect against mastitis causing pathogen Staphylococcus aureus [Pubmed: 30975214]
Journal of Immunology Research Damage-Associated Molecular Patterns and Myeloid-Derived Suppressor Cells in Bronchoalveolar Lavage Fluid in Chronic Obstructive Pulmonary Disease Patients []
Veterinary Research Toll-like receptor 5-mediated IL-17C expression in intestinal epithelial cells enhances epithelial host defense against F4+ ETEC infection [Pubmed: 31221216]
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