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1,25-二羥基維生素D3(DHVD3)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3)

DH-VD3; Calcitriol; Rocaltrol; Calcijex; 1,25-(OH)2D3; 1,25-Dihydroxycholecalciferol; 1,25-Dihydroxy Vitamin D3

  • 1,25-二羥基維生素D3(DHVD3)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 1,25-二羥基維生素D3(DHVD3)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 1,25-二羥基維生素D3(DHVD3)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • CEA467Ge.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1,25-二羥基維生素D3(DHVD3),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1,25-二羥基維生素D3(DHVD3)(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) 95-104 101
EDTA plasma(n=5) 83-95 87
heparin plasma(n=5) 86-98 95

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(jìn)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1,25-二羥基維生素D3(DHVD3),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1,25-二羥基維生素D3(DHVD3)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) 93-101% 81-92% 88-98% 89-96%
EDTA plasma(n=5) 87-102% 92-101% 92-99% 92-101%
heparin plasma(n=5) 78-89% 93-101% 78-103% 96-103%

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。
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DHVD3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加入生物素標(biāo)記的DHVD3類似物和待測抗原(標(biāo)準(zhǔn)品或樣本),待測抗原與生物素標(biāo)記DHVD3類似物對特異性抗體進(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),計算樣品濃度。

相關(guān)產(chǎn)品

編號 適用物種:Pan-species (General,通用) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
CEA467Ge 1,25-二羥基維生素D3(DHVD3)檢測試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA467Ge 1,25-二羥基維生素D3(DHVD3)等多因子檢測試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.

參考文獻(xiàn)

雜志 參考文獻(xiàn)
PLoS One Selective Cytotoxic Action and DNA Damage by Calcitriol-Cu(II) Interaction: Putative Mechanism of Cancer Prevention [PubMed: PMC3785422]
BMC Medicine Calcium-deficiency assessment and biomarker identification by an integrated urinary metabonomics analysis [PubMed: PMC3652781]
Nephrol Dial Transplant. Treatment of established left ventricular hypertrophy with fibroblast growth factor receptor blockade in an animal model of CKD [Pubmed:24875663]
Endocrinology. Vitamin D Prevents Podocyte Injury via Regulation of Macrophage M1/M2 Phenotype in Diabetic Nephropathy Rats [Pubmed:25188527]
Tumor Biology Calcitriol?induced?redox?imbalance and DNA breakage in cells sharing a common metabolic feature of malignancies: Interaction with cellular copper (II) ions leads to the production of reactive oxygen species. [Pubmed:25547436]
International Journal of Life Sciences Research Impact of Curcumin and Flaxseed Oil on Bone Turnover in Glucocorticoid-Injected Female Rats [profile:303311214]
Protective Effect of Soybean and Mushroom against Glucocorticoid-Induced Osteoporosis in Female Rats [:]
Apoptosis Calcitriol–copper interaction leads to non enzymatic, reactive oxygen species mediated DNA breakage and modulation of cellular redox scavengers in hepatocellular … [article:10.1007]
?in vivo Effect of Calcitriol on FGF23 Level in Healthy Adults and its Dependence on Phosphate Level [pubmed:28064234]
Hypertension Research Glucocorticoid exposure induces preeclampsia via dampening 1,25-dihydroxyvitamin D3 [pubmed:29093562]
International?Journal?of?Clinical?and?Experimental?Medicine Combined effect of pulsed electromagnetic fields and narrowband ultraviolet B on bone metabolism in glucocorticoid-treated rats []
Journal of Clinical and Translational Endocrinology Dobesity: relationship between vitamin d deficiency, obesity and sclerostin as a novel biomarker of bone metabolism []
International Journal of Environmental Research and Public Health Alleviating Effect of α-Lipoic Acid and Magnesium on Cadmium-Induced Inflammatory Processes, Oxidative Stress and Bone Metabolism Disorders in Wistar Rats [Pubmed: 31739465]
Ecotoxicology and Environmental Safety Sodium dehydroacetate induces cardiovascular toxicity associated with Ca2+ imbalance in zebrafish []
EXPERIMENTAL AND MOLECULAR MEDICINE Metabolic activities affect femur and lumbar vertebrae remodeling, and anti-resorptive risedronate disturbs femoral cortical bone remodeling [33436949]
Life Sci SWIM domain protein ZSWIM4 is required for JAK2 inhibition resistance in breast cancer [34102191]
Nutrients Effect of Dapagliflozin and Magnesium Supplementation on Renal Magnesium Handling and Magnesium Homeostasis in Metabolic Syndrome [34836340]
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