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補(bǔ)體成分3a(C3a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Complement Component 3a (C3a)

  • 補(bǔ)體成分3a(C3a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 補(bǔ)體成分3a(C3a)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 補(bǔ)體成分3a(C3a)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
  • SEA387Po.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)補(bǔ)體成分3a(C3a)，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補(bǔ)體成分3a(C3a)（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) 97-105 101
EDTA plasma(n=5) 84-101 91
heparin plasma(n=5) 84-98 88

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)加入適量的補(bǔ)體成分3a(C3a)，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補(bǔ)體成分3a(C3a)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) 90-104% 91-99% 85-99% 84-98%
EDTA plasma(n=5) 80-93% 95-103% 93-101% 81-98%
heparin plasma(n=5) 78-97% 78-91% 81-101% 84-101%

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）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

將補(bǔ)體成分3a(C3a)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，其中的補(bǔ)體成分3a(C3a)與連接于固相載體上的抗體結(jié)合，然后加入生物素化的補(bǔ)體成分3a(C3a)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補(bǔ)體成分3a(C3a)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測(cè)定吸光度(O.D.值)，計(jì)算樣品濃度。

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編號(hào) 適用物種：Sus scrofa; Porcine (Pig，豬) 應(yīng)用(僅供研究使用，不用于臨床診斷！)
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MAA387Po21 補(bǔ)體成分3a(C3a)單克隆抗體 WB; IHC; ICC; IP.
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參考文獻(xiàn)

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EXPERIMENTAL EYE RESEARCH Increased ratios of complement factors C3a to C3 in aqueous humor and serum mark glaucoma progression [33493474]
FASEB J C‐reactive protein inhibits C3a/C3aR‐dependent podocyte autophagy in favor of diabetic kidney disease [Pubmed:35503088]
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