產(chǎn)品概述
IL-12,也稱為NK細(xì)胞刺激因子(NKSF)或細(xì)胞毒性淋巴細(xì)胞成熟因子(CLMF),是一種多效性細(xì)胞因子,主要由抗原提呈細(xì)胞(單核/巨噬細(xì)胞、樹(shù)突狀細(xì)胞和B淋巴細(xì)胞)產(chǎn)生。IL-12對(duì)T淋巴細(xì)胞和NK細(xì)胞具有多種效應(yīng),包括刺激細(xì)胞毒作用、增殖、細(xì)胞因子產(chǎn)生以及Th1型應(yīng)答的發(fā)展。IL-12是一個(gè)分子量為 70 kDa(p70)的異二聚體糖蛋白,由一個(gè)40 kDa 亞基(p40)和一個(gè)35 kDa 亞基通過(guò)二硫鍵連接而成。單獨(dú)的p40和p35亞基不具有IL-12活性,但是p40 亞基同源二聚體可以結(jié)合IL-12受體,因此是IL-12拮抗劑。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??谷薎L-12 p40 單抗包被于酶標(biāo)板上,同時(shí)加入標(biāo)本、標(biāo)準(zhǔn)品和生物素化的抗人IL-12 p40抗體,標(biāo)本、標(biāo)準(zhǔn)品中的IL-12 p40 會(huì)與加入于孔中的生物素化的抗人 IL-12 p40 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(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 IL-12 p40,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,IL-12 p40 濃度與OD450值之間呈正相關(guān),可通過(guò)繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中IL-12 p40濃度。
產(chǎn)品性能
1. 靈敏度:最小可測(cè)人IL-12/IL-23 p40達(dá)30pg/ml
2. 重復(fù)性:板內(nèi)、板間變異系數(shù)均<10%。
產(chǎn)品組分及儲(chǔ)存條件
名稱
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96 Tests
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48 Tests
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保存條件
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1. 抗體包被板條 (Microwell Plate)
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8×12
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8×6
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2-8℃
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2.標(biāo)準(zhǔn)品 (Standard)
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2 支(凍干)
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1 支(凍干)
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2-8℃
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3.濃縮生物素化抗體 100× (Biotin-Conjugate)
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2 支 (30μl/支)
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1 支(30μl/支)
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2-8℃
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4.濃縮酶結(jié)合物 100× (Streptavidin-HRP)
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2 支 (60μl/支)
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1 支(60μl/支)
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2-8℃(避光)
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5.試劑稀釋液 (Assay Diluent)
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2 瓶(25ml/瓶)
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1 瓶(25ml/瓶)
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2-8℃
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6.濃縮洗滌液 20× (Wash Buffer Concentrate)
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1 瓶 (30ml/瓶)
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1 瓶(30ml/瓶)
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2-8℃
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7.顯色劑 (Substrate Solution)
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1 瓶(12ml/瓶 )
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1 瓶(6ml/瓶)
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2-8℃(避光)
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8.終止液 (Stop Solution)
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1 瓶(12ml/瓶)
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1 瓶(12ml/瓶)
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2-8℃
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9.封板膠紙
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2 張
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2 張
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|
10.說(shuō)明書(shū)
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1 份
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1 份
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試驗(yàn)所需自備儀器和試劑:
1. 酶標(biāo)儀(450nm檢測(cè)波長(zhǎng)濾光片,570nm或630nm校正波長(zhǎng)濾光片)。
2. 高精度加液器及一次性吸頭:0.5-10μl,2-20μl,20-200μl,200-1000μl。
3. 37℃恒溫箱、蒸餾水或去離子水、微量振蕩器、坐標(biāo)紙。
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