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調節(jié)激活正常T-細胞表達分泌因子(RANTES)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES)

CCL5; SCYA5; SISd; EoCP; Chemokine C-C-Motif Ligand 5; Eosinophil Chemotactic Cytokine; T-cell Specific Protein p288; Small Inducible Cytokine Subfamily A(Cys-Cys)Member 5

  • 調節(jié)激活正常T-細胞表達分泌因子(RANTES)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產品包裝(模擬)
  • 調節(jié)激活正常T-細胞表達分泌因子(RANTES)檢測試劑盒(酶聯(lián)免疫吸附試驗法)產品包裝(模擬)
  • 調節(jié)激活正常T-細胞表達分泌因子(RANTES)檢測試劑盒(酶聯(lián)免疫吸附試驗法)實驗結果圖
  • SEA116Mu.jpg標準曲線圖
  • Certificate通過ISO 9001、ISO 13485質量體系認證

特異性

本試劑盒用于檢測調節(jié)激活正常T-細胞表達分泌因子(RANTES)，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調節(jié)激活正常T-細胞表達分泌因子(RANTES)（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)98-105101
EDTA plasma(n=5)85-9488
heparin plasma(n=5)81-9888

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調節(jié)激活正常T-細胞表達分泌因子(RANTES)，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調節(jié)激活正常T-細胞表達分泌因子(RANTES)含量的測定值與理論值的比率。

樣本1:21:41:81:16
serum(n=5)81-101%88-96%80-102%81-97%
EDTA plasma(n=5)82-95%83-99%87-94%85-93%
heparin plasma(n=5)88-103%83-93%92-99%94-105%

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(shù)。

實驗原理

將調節(jié)激活正常T-細胞表達分泌因子(RANTES)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，其中的調節(jié)激活正常T-細胞表達分泌因子(RANTES)與連接于固相載體上的抗體結合，然后加入生物素化的調節(jié)激活正常T-細胞表達分泌因子(RANTES)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調節(jié)激活正常T-細胞表達分泌因子(RANTES)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。

參考文獻

雜志參考文獻
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