產(chǎn)品概述
人VEGF(Vascular endothelial growth factor)基因編碼8個(gè)外顯子。由于不同剪切形式,產(chǎn)生至少有 4種異構(gòu)體(VEGF121,VEGF165,VEGF189與VEGF206)。VEGF是一種高度保守的蛋白質(zhì),在不同的物種中,都有交叉活力。在人、大鼠或牛中,VEGF有84-94%的序列同源性。在體外,VEGF 是一種具有重要作用的內(nèi)皮細(xì)胞有絲分裂原。在體外培養(yǎng)內(nèi)皮細(xì)胞時(shí),VEGF能激活磷酸脂酶C,誘導(dǎo)胞質(zhì)內(nèi)的自由Ca2+濃度增加。VEGF對(duì)單核細(xì)胞和造骨細(xì)胞也是化學(xué)趨化因子。在體內(nèi),VEGF能誘導(dǎo)血管生成和增加血管滲透性?;谶@些體外與體內(nèi)的活性,VEGF被認(rèn)為在傷口愈合、胚胎發(fā)育、固體瘤的生長(zhǎng)與轉(zhuǎn)移等過(guò)程有重要作用。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薞EGF單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的VEGF會(huì)與單抗結(jié)合,游離的成分被洗去形成免疫復(fù)合物;加入生物素化的抗人VEGF抗體,生物素化抗人VEGF抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的VEGF結(jié)合而形成免疫復(fù)合物,游離的成分被洗去;加入辣根過(guò)氧化物酶標(biāo)記的親合素,親合素與生物素特異性結(jié)合,游離的成分被洗去。加入顯色底物(顯色劑),若反應(yīng)孔中有VEGF,辣根過(guò)氧化物酶會(huì)使無(wú)色的顯色劑變藍(lán),加終止液變黃。在450nm處測(cè)OD值, VEGF濃度與OD450值之間呈正相關(guān),可通過(guò)繪制標(biāo)準(zhǔn)曲線(xiàn)求出標(biāo)本中VEGF濃度。
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