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羥脯氨酸(Hyp)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Hydroxyproline (Hyp)

(2S,4R)-4-Hydroxyproline; L-Hydroxyproline(C5H9O3N)

  • 羥脯氨酸(Hyp)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 羥脯氨酸(Hyp)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 羥脯氨酸(Hyp)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
  • CEA621Ge.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)羥脯氨酸(Hyp),經(jīng)檢測(cè)與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,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羥脯氨酸(Hyp)(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) 85-99 94
EDTA plasma(n=5) 85-98 90
heparin plasma(n=5) 91-105 97

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)加入適量的羥脯氨酸(Hyp),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羥脯氨酸(Hyp)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) 85-104% 97-105% 93-101% 93-105%
EDTA plasma(n=5) 87-95% 97-105% 78-89% 84-101%
heparin plasma(n=5) 83-94% 85-93% 79-88% 78-98%

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)50µL,
    加入50µL檢測(cè)液A(臨用前配制);
    37°C溫育1小時(shí)。
3. 洗板3次;
4. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分鐘;
7. 加終止液50µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

本試劑盒應(yīng)用競(jìng)爭(zhēng)抑制酶聯(lián)免疫分析法測(cè)定標(biāo)本中待測(cè)物質(zhì)水平。將羥脯氨酸(Hyp)單克隆抗體包被微孔板,制成固相載體,往包被抗體的微孔中同時(shí)加入生物素標(biāo)記的抗原和待測(cè)抗原(標(biāo)準(zhǔn)品或樣本),待測(cè)抗原與生物素標(biāo)記抗原對(duì)特異性抗體進(jìn)行競(jìng)爭(zhēng)結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過溫育和徹底洗滌后加入底物TMB顯色。TMB在過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。待測(cè)標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測(cè)物質(zhì)含量呈負(fù)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:Pan-species (General,通用) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
CPA621Ge11 羥脯氨酸(Hyp)牛血清白蛋白偶聯(lián)物 Immunogen; SDS-PAGE; WB.
CPA621Ge21 羥脯氨酸(Hyp)卵白蛋白偶聯(lián)物 Immunogen; SDS-PAGE; WB.
PAA621Ge01 羥脯氨酸(Hyp)多克隆抗體 ELISA, CLIA. / IHC-Fr, ICC, IP (predicted).
CEA621Ge 羥脯氨酸(Hyp)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
MEA621Ge 羥脯氨酸(Hyp)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法,小樣本) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA621Ge 羥脯氨酸(Hyp)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
KSA621Ge11 羥脯氨酸(Hyp)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)
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Canadian Journal of Physiology and Pharmacology Renin–angiotensin system inhibition ameliorates CCl-induced liver fibrosis in mice through the inactivation of nuclear transcription factor kappa B [10.1139:cjpp-2017-0728]
Experimental and Therapeutic Medicine Pirfenidone suppresses bleomycin?induced pulmonary fibrosis and periostin expression in rats [10.3892:etm.2018.6378]
Cell Death & Disease Therapeutic effect of hepatocyte growth factor-overexpressing bone marrow-derived mesenchymal stem cells on CCl 4-induced hepatocirrhosis. []
International wound journal The effects of platelet‐rich‐plasma gel application to the colonic anastomosis in hyperthermic intraperitoneal chemotherapy: An experimental rat model [Pubmed: 31448554]
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY Pantoea agglomerans chronic exposure induces epithelial-mesenchymal transition in human lung epithelial cells and mice lungs [Pubmed: 32146192]
Annals of Surgical Treatment and Research Is platelet-rich plasma improves the anastomotic healing in hyperthermic intraperitoneal chemotherapy with oxaliplatin: an experimental rat study [Pubmed: 32051817]
Arch Med Res Protective Effects of Contractubex? on Stricture Formation after Experimental Corrosive Esophageal Burns in Rats [Pubmed: 32654881]
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Dis Marker Handgrip Strength and Vitamin D as Predictors of Liver Fibrosis and Malnutrition in Chronic Hepatitis C Patients [33884041]
PLoS One Beneficial impact of cathelicidin on hypersensitivity pneumonitis treatment?aIn vivo studies [33999928]
Front Cell Dev Biol Curdione and Schisandrin C Synergistically Reverse Hepatic Fibrosis via Modulating the TGF-β Pathway and Inhibiting Oxidative Stress [34858986]
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