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血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)檢測試劑盒(酶聯(lián)免疫吸附試驗法)

ELISA Kit for A Disintegrin And Metalloproteinase With Thrombospondin 4 (ADAMTS4)

ADMP-1; Aggrecanase-1

  • 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 產(chǎn)品包裝(模擬)
  • 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)檢測試劑盒(酶聯(lián)免疫吸附試驗法) 實驗結(jié)果圖
  • SEK204Hu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認證

特異性

本試劑盒用于檢測血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4),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血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)(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) 96-103 101
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 80-97 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(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)加入適量的血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4),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血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)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) 90-104% 95-104% 97-105% 88-97%
EDTA plasma(n=5) 96-103% 78-91% 89-96% 95-105%
heparin plasma(n=5) 78-95% 83-101% 84-95% 78-99%

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(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;
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

將血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)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,其中的血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)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血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)呈正相關(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。

參考文獻

雜志 參考文獻
Biomedicine & Pharmacotherapy ADAMTS4 level in patients with stable coronary artery disease and acute coronary syndromes [ScienceDirect: S0753332209002017]
PLoS ONE A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches [Plosone: Source]
Anadolu Kardiyol Derg. A novel association between TGFb1 and ADAMTS4 in coronary artery disease: A new potential mechanism in the progression of atherosclerosis and diabetes. [Pubmed:25592103]
Osteoarthritis and Cartilage Lack of anti-inflammatory and anti-catabolic effects on basal inflamed osteoarthritic chondrocytes or synoviocytes by adipose stem cell-conditioned medium [PubMed: 26521751]
Osteoarthritis and Cartilage Parathyroid hormone(1-34) exhibits more comprehensive effects than celecoxib in cartilage metabolism and maintaining subchondral bone micro-architecture in meniscectomized Cavia (Guinea pig )inea pigs [Pubmed:26802547]
Journal of Cellular and Molecular Medicine VIP and CRF reduce ADAMTS expression and function in osteoarthritis synovial fibroblasts [Pubmed:26818776]
Int Wound J. From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases. [Pubmed:26991748]
The American journal of pathology Healthy and Osteoarthritic Synovial Fibroblasts Produce a Disintegrin and Metalloproteinase with Thrombospondin Motifs 4, 5, 7, and 12: Induction by IL-1β and Fibronectin and Contribution to Cartilage Damage. [pubmed:27449198]
International Wound Journal From varices to venous ulceration: the story of chronic venous disease described by metalloproteinases [pubmed:26991748]
American Journal of Translational?Research Diagnostic utility of fluorogenic peptide-conjugated Au nanoparticle probe corroborated by rabbit model of mild cartilage injury and panel of osteoarthritic?… [Pubmed: 30210670]
Journal of Clinical Medicine The Effects of TNF-α Inhibition on the Metabolism of Cartilage: Relationship between KS, HA, HAPLN1 and ADAMTS4, ADAMTS5, TOS and TGF-β1 Plasma?… [Pubmed:35407621]
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