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促腎上皮質(zhì)激素釋放激素(CRF)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Corticotropin Releasing Factor (CRF)

CRH; Corticotropin Releasing Hormone; Corticoliberin

  • 促腎上皮質(zhì)激素釋放激素(CRF)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 促腎上皮質(zhì)激素釋放激素(CRF)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 促腎上皮質(zhì)激素釋放激素(CRF)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • CEA835Hu.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測促腎上皮質(zhì)激素釋放激素(CRF),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促腎上皮質(zhì)激素釋放激素(CRF)(加標(biāo)樣品),重復(fù)測定并計(jì)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) 85-93 89
EDTA plasma(n=5) 91-101 95
heparin plasma(n=5) 93-101 97

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(jìn)行定量檢測,每份樣本連續(xù)測定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低、中、高值定值樣本進(jìn)行定量測定,每個(gè)樣本使用同一試劑盒重復(fù)測定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)加入適量的促腎上皮質(zhì)激素釋放激素(CRF),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促腎上皮質(zhì)激素釋放激素(CRF)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) 86-101% 78-102% 83-99% 97-104%
EDTA plasma(n=5) 83-101% 80-105% 90-98% 80-93%
heparin plasma(n=5) 78-92% 94-101% 85-99% 97-105%

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(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檢測液A(臨用前配制);
    37°C溫育1小時(shí)。
3. 洗板3次;
4. 加檢測溶液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)用競爭抑制酶聯(lián)免疫分析法測定標(biāo)本中待測物質(zhì)水平。將促腎上皮質(zhì)激素釋放激素(CRF)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)記的抗原和待測抗原(標(biāo)準(zhǔn)品或樣本),待測抗原與生物素標(biāo)記抗原對特異性抗體進(jìn)行競爭結(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)過溫育和徹底洗滌后加入底物TMB顯色。TMB在過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),而與樣品中待測物質(zhì)含量呈負(fù)相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(O.D.值),計(jì)算樣品濃度。

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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!)
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參考文獻(xiàn)

雜志 參考文獻(xiàn)
Toxicology Letters Regulation of corticosterone secretion is modified by PFOS exposure at different levels of the hypothalamic–pituitary–adrenal axis in adult male rats [Pubmed:24440345]
Bipolar?Disord. Immune dysfunction in bipolar disorder and suicide risk: is there an association between peripheral corticotropin‐releasing hormone and interleukin‐1β? [Pubmed:24862833]
Peptides Differential stress response in rats subjected to chronic mild stress is accompanied by changes in CRH-family gene expression at the pituitary level [Pubmed:25236411]
Ann Dermatol Acute?Stress-Induced?Changes?in?Follicular?Dermal?Papilla?Cells?and?Mobilization?of?Mast?Cells:Implications?for?Hair?Growth. [pubmed:27746640]
Neuroscience Heat stress-induced neuroinflammation and aberration in monoamine levels in hypothalamus are associated with temperature dysregulation [pubmed:28663093]
Biomed Research International Colonic Transit Time and Gut Peptides in Adult Patients with Slow and Normal Colonic Transit Constipation [pubmed:29082242]
Biochemical Pharmacology Effects of n-3 PUFA enriched and n-3 PUFA deficient diets in na?ve and Aβ-treated female rats [Pubmed:30028991]
Frontiers in Pharmacology Vaccinium bracteatum Leaf Extract Reverses Chronic Restraint Stress-Induced Depression-Like Behavior in Mice: Regulation of Hypothalamic-Pituitary?… [Pubmed:30038568]
Toxicology Letters Perfluorooctane sulfonate (PFOS) can alter the hypothalamic-pituitary-adrenal (HPA) axis activity by modifying CRF1 and glucocorticoid receptors [Pubmed:29807116]
Neuropsychiatric disease and treatment The depressive-like behaviors of chronic unpredictable mild stress-treated mice, ameliorated by Tibetan medicine Zuotai: involvement in the hypothalamic?… [Pubmed:29379286]
BMC?Veterinary?Research Hypothalamus-pituitary-adrenal axis involves in anti-viral ability through regulation of immune response in piglets infected by highly pathogenic porcine?… [Pubmed:29540178]
Bioscience Research Bioscience Research []
FASEB Journal Contribution of DNA methylation in chronic stress–induced cardiac remodeling and arrhythmias in mice [Pubmed: 31431066]
Aging Cell Altered glucocorticoid metabolism represents a feature of macroph‐aging [Pubmed: 32463582]
Endocr J Impairment of CRH in the intestinal mucosal epithelial barrier of pregnant Bama miniature pig induced by restraint stress [33408312]
CRYOBIOLOGY O-GlcNAc/Akt pathway regulates glucose metabolism and reduces apoptosis in liver of piglets with acute cold stress [33651993]
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