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調(diào)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(diào)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(chǎn)品包裝(模擬)
  • 調(diào)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(chǎn)品包裝(模擬)
  • 調(diào)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(jié)果圖
  • SEA116Hu.jpg 標準曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認證

特異性

本試劑盒用于檢測調(diào)節(jié)激活正常T-細胞表達分泌因子(RANTES),經(jīng)檢測與其它相似物質(zhì)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(zhì)交叉反應檢測,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。

回收率

分別于定值血清及血漿樣本中加入一定量的調(diào)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) 97-104 101
EDTA plasma(n=5) 81-101 96
heparin plasma(n=5) 86-95 91

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(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)加入適量的調(diào)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(diào)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 1:2 1:4 1:8 1:16
serum(n=5) 91-103% 89-103% 80-97% 97-105%
EDTA plasma(n=5) 93-101% 78-91% 89-103% 92-105%
heparin plasma(n=5) 78-93% 91-99% 80-102% 78-88%

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(diào)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(diào)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(jié)合,然后加入生物素化的調(diào)節(jié)激活正常T-細胞表達分泌因子(RANTES)抗體,將未結(jié)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(zhuǎn)化成藍色,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調(diào)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。

相關產(chǎn)品

編號 適用物種:Homo sapiens (Human,人) 應用(僅供研究使用,不用于臨床診斷!)
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KSA116Hu01 調(diào)節(jié)激活正常T-細胞表達分泌因子(RANTES)檢測試劑盒DIY材料(酶聯(lián)免疫吸附試驗法) Main materials for "Do It (ELISA Kit) Yourself".

參考文獻

雜志 參考文獻
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Cell Death & Disease CCL5 derived from tumor-associated macrophages promotes prostate cancer stem cells and metastasis via activating β-catenin/STAT3 signaling [Pubmed: 32300100]
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