產(chǎn)品概述
腫瘤壞死因子-α也稱惡病質(zhì)素,腫瘤壞死因子-β也稱淋巴毒素,它們是兩種密切相關(guān)的蛋白質(zhì)(約34%的氨基酸序列同源),這兩種蛋白質(zhì)結(jié)合在相同的細(xì)胞表面受體上并表現(xiàn)出許多共同的生物學(xué)功能。當(dāng)主體遭到感染或惡性腫瘤增殖的時(shí)候作為主要的免疫激活反應(yīng)劑和主要的介導(dǎo)炎癥反應(yīng)劑TNF-α和TNF-β在其中起到了關(guān)鍵的作用。TNF-α由活化巨噬細(xì)胞和其它一些細(xì)胞激活,這些細(xì)胞包括T細(xì)胞、B細(xì)胞、NK細(xì)胞、LAK細(xì)胞、星形膠質(zhì)細(xì)胞、內(nèi)皮細(xì)胞、平滑肌細(xì)胞和一些腫瘤細(xì)胞。大鼠TNF-αcDNA是一個(gè)235個(gè)氨基酸殘基的II型膜蛋白。通過(guò)金屬蛋白酶水解從跨膜區(qū)C端釋放出一個(gè)由156個(gè)氨基酸組成的可溶TNF-α(sTNF-α)。
產(chǎn)品原理
本實(shí)驗(yàn)采用雙抗體夾心ELISA法??勾笫骉NF-α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TNF-α抗體,標(biāo)本、標(biāo)準(zhǔn)品中的TNF-α?xí)c加入于孔中的生物素化的抗大鼠TNF-α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TNF-α,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,TNF-α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TNF-α濃度。
產(chǎn)品性能
1. 靈敏度:最小可測(cè)大鼠TNF-α達(dá)15pg/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. 微孔板振蕩器、蒸餾水或去離子水、坐標(biāo)紙。
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