產(chǎn)品概述
人的瘦素蛋白是一個16KDa含有146個氨基酸殘基的沒有糖基化修飾的多肽,一開始使用基因圖譜技術(shù)從肥胖老鼠的基因中定位到一個突變,這個基因就是編碼瘦素蛋白的基因。由于它能夠降低脂肪含量人們把它命名為瘦素(OB),它在不同種屬中有很高的保守性,成熟人的瘦素蛋白與小鼠和大鼠分別有87%和84%的同源性。瘦素通常被稱為OB蛋白質(zhì),這種蛋白質(zhì)是ob基因(obese gene)的產(chǎn)物。人的瘦素被發(fā)現(xiàn)無論在小鼠或者大鼠的系統(tǒng)中都是有活性的。瘦素的表達(dá)是非常有限的,僅在脂肪細(xì)胞中表達(dá)。瘦素蛋白主要被認(rèn)為調(diào)控降低食欲和能量消耗。當(dāng)脂肪細(xì)胞攝入大量卡路里的時候,它的大小會變大,從而釋放出瘦素蛋白。大的脂肪細(xì)胞說明短期內(nèi)不需要能量攝入,循環(huán)的瘦素蛋白給控制食欲和代謝速率的下丘腦傳達(dá)了抑制食欲的信號。盡管在人類和小鼠中瘦素的循環(huán)水平和體重呈現(xiàn)正相關(guān)性,但是許多因子在大腦和周邊組織中與體重減輕和代謝也有很大關(guān)系。除了它在脂肪代謝中起的作用之外,瘦素在造血和腎上腺皮質(zhì)功能中也發(fā)揮重要作用。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薒eptin單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的Leptin會與單抗結(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Leptin抗體,生物素化抗人Leptin抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的Leptin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Leptin,辣根過氧化物酶會使無色的顯色劑變藍(lán),加終止液變黃。在450nm處測OD值, Leptin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Leptin濃度。
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