Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
ELISA Kit for Collagen Type IV (COL4)
Type-IV Collagen
- 編號SEA180Rb
- 物種Oryctolagus cuniculus (Rabbit,兔) 相同的名稱,不同的物種。
- 實驗方法雙抗夾心
- 反應時長3h
- 檢測范圍78-5,000pg/mL
- 靈敏度最小可檢測劑量小于等于31pg/mL.
- 樣本類型Serum, plasma and other biological fluids
- 下載 英文說明書 中文說明書
- 規(guī)格 48T96T 96T*5 96T*10 96T*100
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特異性
本試劑盒用于檢測Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)(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) | 88-96 | 92 |
EDTA plasma(n=5) | 83-93 | 87 |
heparin plasma(n=5) | 88-101 | 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(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Ⅳ型膠原(COL4),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Ⅳ型膠原(COL4)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) | 80-92% | 90-102% | 93-101% | 87-102% |
EDTA plasma(n=5) | 98-105% | 83-99% | 80-99% | 86-97% |
heparin plasma(n=5) | 91-101% | 81-101% | 85-95% | 95-104% |
穩(wěn)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(huán)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Ⅳ型膠原(COL4)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Ⅳ型膠原(COL4)與連接于固相載體上的抗體結合,然后加入生物素化的Ⅳ型膠原(COL4)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Ⅳ型膠原(COL4)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:Oryctolagus cuniculus (Rabbit,兔) | 應用(僅供研究使用,不用于臨床診斷!) |
SEA180Rb | Ⅳ型膠原(COL4)檢測試劑盒(酶聯(lián)免疫吸附試驗法) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMA180Rb | Ⅳ型膠原(COL4)等多因子檢測試劑盒(流式熒光發(fā)光法) | FLIA Kit for Antigen Detection. |
參考文獻
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