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免疫球蛋白M(IgM)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Immunoglobulin M (IgM)

IGHM; Immunoglobulin Heavy Constant Mu; Ig mu chain C region

  • 免疫球蛋白M(IgM)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 免疫球蛋白M(IgM)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 免疫球蛋白M(IgM)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 實驗結(jié)果圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測免疫球蛋白M(IgM),經(jīng)檢測與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,不可能完成對所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測,因此本試劑盒有可能與未經(jīng)檢測的其它物質(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免疫球蛋白M(IgM)(加標(biāo)樣品),重復(fù)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) 96-104 99
EDTA plasma(n=5) 82-92 88
heparin plasma(n=5) 89-101 94

精密度

精密度用樣品測定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對低、中、高值定值樣本進(jìn)行定量檢測,每份樣本連續(xù)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(jìn)行定量測定,每個樣本使用同一試劑盒重復(fù)測定8次,分別計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)加入適量的免疫球蛋白M(IgM),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免疫球蛋白M(IgM)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) 87-101% 94-101% 89-96% 96-103%
EDTA plasma(n=5) 85-93% 86-98% 78-97% 90-99%
heparin plasma(n=5) 84-101% 89-101% 92-101% 81-95%

穩(wěn)定性

經(jīng)測定,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%。
為減小外部因素對試劑盒破壞前后檢測值的影響,實驗室的環(huán)境條件需盡量保持一致,尤其是實驗室內(nèi)溫度、濕度及溫育條件。其次由同一實驗員來進(jìn)行操作可減少人為誤差。

實驗流程

1. 實驗前標(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檢測液A(臨用前配制);
    37°C溫育1小時。
3. 洗板3次;
4. 加檢測溶液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)。

實驗原理

本試劑盒應(yīng)用競爭抑制酶聯(lián)免疫分析法測定標(biāo)本中待測物質(zhì)水平。將免疫球蛋白M(IgM)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加入生物素標(biāo)記的抗原和待測抗原(標(biāo)準(zhǔn)品或樣本),待測抗原與生物素標(biāo)記抗原對特異性抗體進(jìn)行競爭結(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),而與樣品中待測物質(zhì)含量呈負(fù)相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。

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參考文獻(xiàn)

雜志 參考文獻(xiàn)
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Food and Agricultural Immunology Effects of and on growth performance, immune function and volatile fatty acid level of caecal digesta in broilers [10.1080:09540105.2018.1457013]
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International Journal of Biological Macromolecules Effects of polysaccharides from Yingshan Yunwu tea on meat quality, immune status and intestinal microflora in chickens [Pubmed: 32224178]
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BIOLOGICAL TRACE ELEMENT RESEARCH Association Between Serum Selenium Level and Subclinical Mastitis in Dairy Cattle [Pubmed: 32583225]
IMMUNOSTIMULATORY EFFECT OF MORINGA OLEIFERA LEAVES EXTRACT ON CYCLOPHOSPHAMIDE-INDUCED IMMUNOSUPPRESSION IN MICE []
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