食欲素A(OXA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)
ELISA Kit for Orexin A (OXA)
Hypocretin-1
- 編號(hào)CEA607Mu
- 物種Mus musculus (Mouse,小鼠) 相同的名稱,不同的物種。
- 實(shí)驗(yàn)方法競(jìng)爭(zhēng)抑制
- 反應(yīng)時(shí)長(zhǎng)2h
- 檢測(cè)范圍12.35-1,000pg/mL
- 靈敏度最小可檢測(cè)劑量小于等于5.05pg/mL.
- 樣本類型serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- 下載 英文說(shuō)明書(shū) 中文說(shuō)明書(shū)
- 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(cè)食欲素A(OXA),經(jīng)檢測(cè)與其它相似物質(zhì)無(wú)明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來(lái)源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)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食欲素A(OXA)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。
樣本 | 回收率范圍(%) | 平均回收率(%) |
serum(n=5) | 83-93 | 90 |
EDTA plasma(n=5) | 89-103 | 94 |
heparin plasma(n=5) | 84-92 | 88 |
精密度
精密度用樣品測(cè)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(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)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)加入適量的食欲素A(OXA),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中食欲素A(OXA)含量的測(cè)定值與理論值的比率。
樣本 | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 90-97% | 87-101% | 81-89% | 82-91% |
EDTA plasma(n=5) | 88-103% | 81-99% | 97-104% | 88-103% |
heparin plasma(n=5) | 87-103% | 87-101% | 82-101% | 98-105% |
穩(wěn)定性
經(jīng)測(cè)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(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來(lái)進(jìn)行操作可減少人為誤差。
實(shí)驗(yàn)流程
1. 實(shí)驗(yàn)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)50µL,
加入50µL檢測(cè)液A(臨用前配制);
37°C溫育1小時(shí)。
3. 洗板3次;
4. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分鐘;
7. 加終止液50µL,立即450nm讀數(shù)。
實(shí)驗(yàn)原理
本試劑盒應(yīng)用競(jìng)爭(zhēng)抑制酶聯(lián)免疫分析法測(cè)定標(biāo)本中待測(cè)物質(zhì)水平。將食欲素A(OXA)單克隆抗體包被微孔板,制成固相載體,往包被抗體的微孔中同時(shí)加入生物素標(biāo)記的抗原和待測(cè)抗原(標(biāo)準(zhǔn)品或樣本),待測(cè)抗原與生物素標(biāo)記抗原對(duì)特異性抗體進(jìn)行競(jìng)爭(zhēng)結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過(guò)溫育和徹底洗滌后加入底物TMB顯色。TMB在過(guò)氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。待測(cè)標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測(cè)物質(zhì)含量呈負(fù)相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。
贈(zèng)品
相關(guān)產(chǎn)品
編號(hào) | 適用物種:Mus musculus (Mouse,小鼠) | 應(yīng)用(僅供研究使用,不用于臨床診斷!) |
RPA607Mu01 | 食欲素A(OXA)重組蛋白 | Positive Control; Immunogen; SDS-PAGE; WB. |
PAA607Mu01 | 食欲素A(OXA)多克隆抗體 | WB; IHC; ICC; IP. |
CEA607Mu | 食欲素A(OXA)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) | Enzyme-linked immunosorbent assay for Antigen Detection. |
參考文獻(xiàn)
雜志 | 參考文獻(xiàn) |
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Mol Med Rep | Wake-promoting actions of median nerve stimulation in TBI-induced coma: An investigation of orexin-A and orexin receptor 1 in the hypothalamic region [PubMed: 26059340] |
Mol Neurobiol | Application of the Co-culture Membrane System Pointed to a Protective Role of Catestatin on Hippocampal Plus Hypothalamic Neurons Exposed to Oxygen and Glucose Deprivation [article:10.1007] |
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Journal of Translational Medicine | Recurrent circadian fasting (RCF) improves blood pressure, biomarkers of cardiometabolic risk and regulates inflammation in men [Pubmed: 31426866] |
iraqi journal of science | Relationship Between Orexin-A and Insulin Resistance in Patients with Type 2 Diabetes Mellitus [] |
Brain Sci | Changes in the Orexin System in Rats Exhibiting Learned Helplessness Behaviors [34942932] |