產(chǎn)品概述
血管內(nèi)皮生長(zhǎng)因子(Vascular endothelial growth factor)又稱血管通透因子(VPF),是生長(zhǎng)因子VEGF/PDGF家族的一個(gè)成員。VEGF基因編碼8個(gè)外顯子。由于不同剪切形式,產(chǎn)生3種同型異構(gòu)體(VEGF120,VEGF164,VEGF188)。大鼠VEGF164與小鼠和人的氨基酸序列的同源性分別是98%和90%。多種細(xì)胞,包括骨骼和心肌,肝細(xì)胞,造骨細(xì)胞,嗜中性粒細(xì)胞,巨噬細(xì)胞,角朊細(xì)胞,CD34+ 細(xì)胞都能表達(dá)VEGF。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??勾笫骎EGF單抗包被于酶標(biāo)板上,同時(shí)加入標(biāo)本、標(biāo)準(zhǔn)品和生物素化的抗大鼠VEGF抗體,標(biāo)本、標(biāo)準(zhǔn)品中的VEGF會(huì)與加入于孔中的生物素化的抗大鼠抗體及包被于酶標(biāo)板上的單抗結(jié)合,形成免疫復(fù)合物,游離的成分被洗去;加入辣根過氧化物酶標(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,辣根過氧化物酶會(huì)使無色的顯色劑變藍(lán),加終止液變黃。在450nm處測(cè)OD值, IL-1β濃度與OD450值之間呈正相關(guān),可通過繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中VEGF濃度。
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