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血管內(nèi)皮生長因子A(VEGFA)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
編號SEA143Ca
物種Canis familiaris; Canine (Dog，犬) 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15.6-1,000pg/mL
靈敏度最小可檢測劑量小于等于5.6pg/mL.
樣本類型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
規(guī)格48T96T 96T*5 96T*10 96T*100
價格￥ 2008
產(chǎn)品包裝圖模擬
特異性
本試劑盒用于檢測血管內(nèi)皮生長因子A(VEGFA)，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管內(nèi)皮生長因子A(VEGFA)（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) | 88-95 | 92 |
| EDTA plasma(n=5) | 83-93 | 88 |
| heparin plasma(n=5) | 88-102 | 91 |
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)加入適量的血管內(nèi)皮生長因子A(VEGFA)，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管內(nèi)皮生長因子A(VEGFA)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) | 79-102% | 79-103% | 94-103% | 79-97% |
| EDTA plasma(n=5) | 89-98% | 93-104% | 90-98% | 83-98% |
| heparin plasma(n=5) | 84-91% | 78-88% | 78-98% | 87-105% |
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
將血管內(nèi)皮生長因子A(VEGFA)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，其中的血管內(nèi)皮生長因子A(VEGFA)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管內(nèi)皮生長因子A(VEGFA)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管內(nèi)皮生長因子A(VEGFA)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。
參考文獻
| 雜志 | 參考文獻 |
| The Journal of Biological Chemistry | Intertissue flow of GSH as a tumor growth promoting mechanism: interleukin 6 induces GSH release from hepatocytes in metastatic B16 melanoma-bearing mice [Jcb: 196261] |
| Turkish Journal of Veterinary and Animal Sciences | Can the vascular endothelial growth factor (VEGF) and total estradiol 17β be used as a marker of equine oocyte maturation? [Tubitak: source] |
| Regulatory Peptides | The correlation between resting serum leptin and serum angiogenic indices at rest and after submaximal exercise [ScienceDirect: S016701151100156X] |
| Acta Biomaterialia | Multiple release of polyplexes of plasmids VEGF and bFGF from electrospun fibrous scaffolds towards regeneration of mature blood vessels [PubMed: 22484697] |
| Oncogene | Elevated PLGF contributes to small-cell lung cancer brain metastasis [PubMed: 22797069] |
| International Journal of Nephrology | Impairment and Differential Expression of PR3 and MPO on Peripheral Myelomonocytic Cells with Endothelial Properties in Granulomatosis with Polyangiitis [PubMed: 22792461] |
| Zahedan Journal of Research in Medical Sciences | The Effect of Acute Sub-Maximal Endurance Exercise on Serum Angiogenic Indices in Sedentary Men [Zjrms: Source] |
| Nanoscale Research Letters | Silver nanoparticles administered to chicken affect VEGFA and FGF2 gene expression in breast muscle and heart. [PubMed: 22827927] |
| BMC cell biology | BMSCs reduce rat granulosa cell apoptosis induced by cisplatin and perimenopause [PubMed: PMC3640998] |
| PLoS One. | Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing [PubMed: PMC3378547] |
| Oncology Reports | A bone-seeking clone exhibits different biological properties from the ACHN parental human renal cell carcinoma in vivo and in vitro [PubMed: PMC3583590] |
| The Journal of Biological Chemistry | Intertissue Flow of Glutathione (GSH) as a Tumor Growth-promoting Mechanism: INTERLEUKIN 6 INDUCES GSH RELEASE FROM HEPATOCYTES IN METASTATIC B16 MELANOMA-BEARING MICE [PubMed: PMC3091180] |
| International Journal of Molecular Sciences | Nano-Nutrition of Chicken Embryos. The Effect of in Ovo Administration of Diamond Nanoparticles and L-Glutamine on Molecular Responses in Chicken Embryo Pectoral Muscles [NCBI:PMC3856104] |
| Archives of Animal Nutrition | Nano-nutrition of chicken embryos. The effect of silver nanoparticles and ATP on expression of chosen genes involved in myogenesis [Pubmed: 23952606] |
| Angiogenesis | Prodrug of green tea epigallocatechin-3-gallate (Pro-EGCG) as a potent anti-angiogenesis agent for endometriosis in mice [Pubmed: 22948799] |
| Experimental Eye Research | Hyperglycemia promotes vasculogenesis in choroidal neovascularization in diabetic mice by stimulating VEGF and SDF-1 expression in retinal pigment epithelial cells [Pubmed: 24780853] |
| Tumor Biology | Biglycan up-regulated vascular endothelial growth factor (VEGF) expression and promoted angiogenesis in colon cancer [Pubmed:25371074] |
| Archives of Animal Nutrition | Effect of silver nanoparticles and hydroxyproline, administered in ovo, on the development of blood vessels and cartilage collagen structure in chicken embryos [Pubmed:25530495] |
| Stem Cells International | Matrix Metalloproteinase 9 Secreted by Hypoxia Cardiac Fibroblasts Triggers Cardiac Stem Cell Migration In Vitro [Pubmed:25767513] |
| Physiology and Pharmacology | A Study on the Serum Levels of Angiogenic Factors in Response to Acute Long-term Submaximal Exercise in Sedentary Men [Phypha:Source] |
| Int J Clin Exp Med. | The effects of self-assembling peptide RADA16 hydrogel on malignant phenotype of human hepatocellular carcinoma cell [PubMed: 26628972] |
| Mol Med Rep | Application of recombinant peroxisome proliferator [PubMed: 26676891] |
| Journal of Immunotherapy | Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity [PubMed: 25962106] |
| shock | Angiogenesis inhibitor endostatin protects mice with sepsis from multiple organ dysfunction syndrome [PubMed: 26125086] |
| Clinical Science | Pigment epithelium-derived factor regulates microvascular permeability through adipose triglyceride lipase in sepsis [PubMed: 25700221] |
| Histol Histopathol | Estrogen-deficient osteoporosis enhances the recruitment and activity of osteoclasts by breast cancer cells [PubMed: 26254457] |
| International Journal of Oncology | Silencing of Livin inhibits tumorigenesis and metastasis via VEGF and MMPs pathway in lung cancer [PubMed: 26094984] |
| Acta Biomater | Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: an in vitro osteoblast-osteoclast-endothelial cell co-culture study [PubMed: 26689470] |
| JBiSE | Healing Mechanism and Osteogenic Capacity of Bovine Bone Mineral—Human Amniotic Mesenchymal Stem Celland Autogenous Bone Graft in Critical Size Mandibular Defect [Journal: 60629] |
血管內(nèi)皮生長因子A(VEGFA)檢測試劑盒(酶聯(lián)免疫吸附試驗法)
編號SEA143Ca
物種Canis familiaris; Canine (Dog，犬) 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15.6-1,000pg/mL
靈敏度最小可檢測劑量小于等于5.6pg/mL.
樣本類型Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
規(guī)格48T96T 96T*5 96T*10 96T*100
價格￥ 2008
產(chǎn)品包裝圖模擬
特異性
本試劑盒用于檢測血管內(nèi)皮生長因子A(VEGFA)，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管內(nèi)皮生長因子A(VEGFA)（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) | 88-95 | 92 |
| EDTA plasma(n=5) | 83-93 | 88 |
| heparin plasma(n=5) | 88-102 | 91 |
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)加入適量的血管內(nèi)皮生長因子A(VEGFA)，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管內(nèi)皮生長因子A(VEGFA)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) | 79-102% | 79-103% | 94-103% | 79-97% |
| EDTA plasma(n=5) | 89-98% | 93-104% | 90-98% | 83-98% |
| heparin plasma(n=5) | 84-91% | 78-88% | 78-98% | 87-105% |
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
將血管內(nèi)皮生長因子A(VEGFA)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，其中的血管內(nèi)皮生長因子A(VEGFA)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管內(nèi)皮生長因子A(VEGFA)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管內(nèi)皮生長因子A(VEGFA)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。
參考文獻
| 雜志 | 參考文獻 |
| The Journal of Biological Chemistry | Intertissue flow of GSH as a tumor growth promoting mechanism: interleukin 6 induces GSH release from hepatocytes in metastatic B16 melanoma-bearing mice [Jcb: 196261] |
| Turkish Journal of Veterinary and Animal Sciences | Can the vascular endothelial growth factor (VEGF) and total estradiol 17β be used as a marker of equine oocyte maturation? [Tubitak: source] |
| Regulatory Peptides | The correlation between resting serum leptin and serum angiogenic indices at rest and after submaximal exercise [ScienceDirect: S016701151100156X] |
| Acta Biomaterialia | Multiple release of polyplexes of plasmids VEGF and bFGF from electrospun fibrous scaffolds towards regeneration of mature blood vessels [PubMed: 22484697] |
| Oncogene | Elevated PLGF contributes to small-cell lung cancer brain metastasis [PubMed: 22797069] |
| International Journal of Nephrology | Impairment and Differential Expression of PR3 and MPO on Peripheral Myelomonocytic Cells with Endothelial Properties in Granulomatosis with Polyangiitis [PubMed: 22792461] |
| Zahedan Journal of Research in Medical Sciences | The Effect of Acute Sub-Maximal Endurance Exercise on Serum Angiogenic Indices in Sedentary Men [Zjrms: Source] |
| Nanoscale Research Letters | Silver nanoparticles administered to chicken affect VEGFA and FGF2 gene expression in breast muscle and heart. [PubMed: 22827927] |
| BMC cell biology | BMSCs reduce rat granulosa cell apoptosis induced by cisplatin and perimenopause [PubMed: PMC3640998] |
| PLoS One. | Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing [PubMed: PMC3378547] |
| Oncology Reports | A bone-seeking clone exhibits different biological properties from the ACHN parental human renal cell carcinoma in vivo and in vitro [PubMed: PMC3583590] |
| The Journal of Biological Chemistry | Intertissue Flow of Glutathione (GSH) as a Tumor Growth-promoting Mechanism: INTERLEUKIN 6 INDUCES GSH RELEASE FROM HEPATOCYTES IN METASTATIC B16 MELANOMA-BEARING MICE [PubMed: PMC3091180] |
| International Journal of Molecular Sciences | Nano-Nutrition of Chicken Embryos. The Effect of in Ovo Administration of Diamond Nanoparticles and L-Glutamine on Molecular Responses in Chicken Embryo Pectoral Muscles [NCBI:PMC3856104] |
| Archives of Animal Nutrition | Nano-nutrition of chicken embryos. The effect of silver nanoparticles and ATP on expression of chosen genes involved in myogenesis [Pubmed: 23952606] |
| Angiogenesis | Prodrug of green tea epigallocatechin-3-gallate (Pro-EGCG) as a potent anti-angiogenesis agent for endometriosis in mice [Pubmed: 22948799] |
| Experimental Eye Research | Hyperglycemia promotes vasculogenesis in choroidal neovascularization in diabetic mice by stimulating VEGF and SDF-1 expression in retinal pigment epithelial cells [Pubmed: 24780853] |
| Tumor Biology | Biglycan up-regulated vascular endothelial growth factor (VEGF) expression and promoted angiogenesis in colon cancer [Pubmed:25371074] |
| Archives of Animal Nutrition | Effect of silver nanoparticles and hydroxyproline, administered in ovo, on the development of blood vessels and cartilage collagen structure in chicken embryos [Pubmed:25530495] |
| Stem Cells International | Matrix Metalloproteinase 9 Secreted by Hypoxia Cardiac Fibroblasts Triggers Cardiac Stem Cell Migration In Vitro [Pubmed:25767513] |
| Physiology and Pharmacology | A Study on the Serum Levels of Angiogenic Factors in Response to Acute Long-term Submaximal Exercise in Sedentary Men [Phypha:Source] |
| Int J Clin Exp Med. | The effects of self-assembling peptide RADA16 hydrogel on malignant phenotype of human hepatocellular carcinoma cell [PubMed: 26628972] |
| Mol Med Rep | Application of recombinant peroxisome proliferator [PubMed: 26676891] |
| Journal of Immunotherapy | Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity [PubMed: 25962106] |
| shock | Angiogenesis inhibitor endostatin protects mice with sepsis from multiple organ dysfunction syndrome [PubMed: 26125086] |
| Clinical Science | Pigment epithelium-derived factor regulates microvascular permeability through adipose triglyceride lipase in sepsis [PubMed: 25700221] |
| Histol Histopathol | Estrogen-deficient osteoporosis enhances the recruitment and activity of osteoclasts by breast cancer cells [PubMed: 26254457] |
| International Journal of Oncology | Silencing of Livin inhibits tumorigenesis and metastasis via VEGF and MMPs pathway in lung cancer [PubMed: 26094984] |
| Acta Biomater | Antioxidant and bone repair properties of quercetin-functionalized hydroxyapatite: an in vitro osteoblast-osteoclast-endothelial cell co-culture study [PubMed: 26689470] |
| JBiSE | Healing Mechanism and Osteogenic Capacity of Bovine Bone Mineral—Human Amniotic Mesenchymal Stem Celland Autogenous Bone Graft in Critical Size Mandibular Defect [Journal: 60629] |
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