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過(guò)氧化物酶體增殖物激活受體γ(PPARg)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Peroxisome Proliferator Activated Receptor Gamma (PPARg)

PPAR-G; PPARG1; PPARG2; NR1C3; Glitazone Receptor; Nuclear Receptor Subfamily 1 Group C Member 3

  • 過(guò)氧化物酶體增殖物激活受體γ(PPARg)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
  • 過(guò)氧化物酶體增殖物激活受體γ(PPARg)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)產(chǎn)品包裝(模擬)
  • 過(guò)氧化物酶體增殖物激活受體γ(PPARg)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)實(shí)驗(yàn)結(jié)果圖
  • SEA886Rb.jpg標(biāo)準(zhǔn)曲線圖
  • Certificate通過(guò)ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)過(guò)氧化物酶體增殖物激活受體γ(PPARg),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過(guò)氧化物酶體增殖物激活受體γ(PPARg)(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)97-104101
EDTA plasma(n=5)79-10288
heparin plasma(n=5)87-9590

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)加入適量的過(guò)氧化物酶體增殖物激活受體γ(PPARg),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過(guò)氧化物酶體增殖物激活受體γ(PPARg)含量的測(cè)定值與理論值的比率。

樣本1:21:41:81:16
serum(n=5)82-101%95-103%80-88%93-104%
EDTA plasma(n=5)88-101%84-96%87-101%80-89%
heparin plasma(n=5)97-105%94-102%87-101%88-102%

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)100µL,37°C孵育1小時(shí);
3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
4. 洗板3次;
5. 加檢測(cè)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(shí)驗(yàn)原理

將過(guò)氧化物酶體增殖物激活受體γ(PPARg)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,其中的過(guò)氧化物酶體增殖物激活受體γ(PPARg)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的過(guò)氧化物酶體增殖物激活受體γ(PPARg)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(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。顏色的深淺和樣品中的過(guò)氧化物酶體增殖物激活受體γ(PPARg)呈正相關(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。

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參考文獻(xiàn)

雜志參考文獻(xiàn)
Neuroscience LettersElevated levels of PPAR-gamma in the cerebrospinal fluid of patients with multiple sclerosis[Pubmed: 24021801]
J Cell Biochem.Nonivamide enhances miRNA let‐7d expression and decreases adipogenesis PPARγ expression in 3T3‐L1 cells[Pubmed:25704235]
Osteoarthritis CartilageEstablishment of a rabbit model to study the influence of advanced glycation end products accumulation on osteoarthritis and the protective effect of pioglitazone[PubMed: 26321377]
J Clin Diagn Res.Evaluation of Protein Kinase Cβ and PPARγ Activity in Diabetic Rats Supplemented with Momordica charantia[pmc:PMC4866090]
Journal of Clinical&Diagnostic ResearchEvaluation?of?Protein?Kinase?Cβ and PPARγ?Activity?in?Diabetic?RatsSupplemented?with?Momordica?charantia.[pubmed:27190792]
Osteoarthritis?and?CartilageEstablishment?of a?rabbit?model?to?study?the?influence?of?advanced?glycation?end?productsaccumulation?on?osteoarthritis?and the?protective?effect?of pioglitazone.[pubmed:26321377]
Neuroscience LettersUnlike PPARgamma, neither other PPARs nor PGC-1alpha is elevated in the cerebrospinal fluid of patients with multiple sclerosis[pubmed:28483651]
European?Journal?of?ImmunologyEngulfment of Hb‐activated platelets differentiates monocytes into pro‐inflammatory macrophages in PNH patients[Pubmed:29677388]
Histochemistry and?Cell BiologySimpson–Golabi–Behmel syndrome human adipocytes reveal a changing phenotype throughout differentiation[Pubmed:29574488]
molecular and cellular biochemistryMaternal omega-3 fatty acids and vitamin E improve placental angiogenesis in late-onset but not early-onset preeclampsia[Pubmed: 31420792]
Nutrients.?Hyperglycemia Changes Expression of Key Adipogenesis Markers (C/EBPα and PPAR?) and Morphology of Differentiating Human Visceral Adipocytes[Pubmed: 31398873]
journal of biochemical and molecular toxicologyIndomethacin and juglone inhibit inflammatory molecules to induce apoptosis in colon cancer cells[Pubmed: 31916655]
J Cosmet DermatolIn?\vitro effect of pine bark extract on melanin synthesis, tyrosinase activity, production of endothelin?\1 and PPAR in cultured melanocytes exposed to Ultraviolet??-[33960120]
American Chemical SocietyTRPA1 Agonist Cinnamaldehyde Decreases Adipogenesis in 3T3-L1 Cells More Potently than the Non-agonist Structural Analog Cinnamyl Isobutyrate[33403292]
PLoS OneQuantitative real-time measurement of endothelin-1-induced contraction in single non-activated hepatic stellate cells[34343209]
Prostaglandins Leukot Essent Fatty AcidsMaternal Vitamin D Deficiency Reduces Docosahexaenoic Acid, Placental Growth Factor and Peroxisome Proliferator Activated Receptor Gamma levels in the Pup?…[34768025]
Food and Chemical ToxicologyMelatonin attenuates cisplatin-induced acute kidney injury in mice: Involvement of PPARα and fatty acid oxidation[Pubmed:35367536]