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防御素β2(DEFb2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)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(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 防御素β2(DEFb2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 防御素β2(DEFb2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • SEA072Hu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過(guò)ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)防御素β2(DEFb2),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

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(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 84-101 95
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 85-99 95

精密度

精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)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(cè)樣本,線性范圍即為稀釋后樣本中防御素β2(DEFb2)含量的測(cè)定值與理論值的比率。

樣本 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%

穩(wěn)定性

經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。

實(shí)驗(yàn)流程

1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
4. 洗板3次;
5. 加檢測(cè)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(shù)。

實(shí)驗(yàn)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(biāo)準(zhǔn)品或標(biāo)本,其中的防御素β2(DEFb2)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的防御素β2(DEFb2)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)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(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

相關(guān)產(chǎn)品

編號(hào) 適用物種:Homo sapiens (Human,人) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
APA072Hu01 防御素β2(DEFb2)活性蛋白 Cell?culture;?Activity?Assays.
RPA072Hu01 防御素β2(DEFb2)重組蛋白 Positive Control; Immunogen; SDS-PAGE; WB.
PAA072Hu01 防御素β2(DEFb2)多克隆抗體 WB
LAA072Hu71 防御素β2(DEFb2)多克隆抗體(生物素標(biāo)記) WB; IHC; ICC.
MAA072Hu22 防御素β2(DEFb2)單克隆抗體 WB; IHC; ICC; IP.
SEA072Hu 防御素β2(DEFb2)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA072Hu 防御素β2(DEFb2)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
KSA072Hu01 防御素β2(DEFb2)檢測(cè)試劑盒DIY材料(酶聯(lián)免疫吸附試驗(yàn)法) Main materials for "Do It (ELISA Kit) Yourself".

參考文獻(xiàn)

雜志 參考文獻(xiàn)
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(xué)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]
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]
Medicine 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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