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
- 編號SEK204Hu
- 物種Homo sapiens (Human,人) 相同的名稱,不同的物種。
- 實驗方法雙抗夾心
- 反應(yīng)時長3h
- 檢測范圍0.312-20ng/mL
- 靈敏度最小可檢測劑量小于等于0.115ng/mL.
- 樣本類型Serum, plasma, tissue homogenates 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準品或標本,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)。用酶標儀在450nm波長下測定吸光度(O.D.值),計算樣品濃度。
增值服務(wù)
相關(guān)產(chǎn)品
編號 | 適用物種:Homo sapiens (Human,人) | 應(yīng)用(僅供研究使用,不用于臨床診斷!) |
RPK204Hu01 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)重組蛋白 | Positive Control; Immunogen; SDS-PAGE; WB. |
PAK204Hu01 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)多克隆抗體 | WB; IHC; ICC; IP. |
LAK204Hu71 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)多克隆抗體(生物素標記) | WB; IHC; ICC. |
LAK204Hu81 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)多克隆抗體(異硫氰酸熒光素標記) | WB; IHC; ICC; IF. |
MAK204Hu21 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)單克隆抗體 | WB; IHC; ICC; IP. |
SEK204Hu | 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) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMK204Hu | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)等多因子檢測試劑盒(流式熒光發(fā)光法) | FLIA Kit for Antigen Detection. |
KSK204Hu01 | 血小板反應(yīng)蛋白解整合素金屬肽酶4(ADAMTS4)檢測試劑盒DIY材料(酶聯(lián)免疫吸附試驗法) | 自己動手制作ELISA試劑盒的主要材料 |
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] |