產(chǎn)品概述
RANTES(Regulated upon Activation, Normal T cell Expressed andpresumably Secreted),也稱為CCL5,是趨化因子“CC”亞家族的一個成員,通過趨化白細(xì)胞以及調(diào)節(jié)這些細(xì)胞的功能從而在炎癥免疫應(yīng)答中發(fā)揮其主要作用。人RANTES cDNA編碼一個高度堿性的含91個氨基酸殘基的多肽前體,該多肽前體含一個23個氨基酸殘基的疏水性信號肽,切除該信號肽后產(chǎn)生含68個氨基酸殘基的成熟蛋白。在推斷的氨基酸水平上,人RANTES和小鼠RANTES的同源性大約為85%。 RANTES對多種不同種類的細(xì)胞都具有強(qiáng)力趨化作用,這些細(xì)胞包括未刺激的CD4+/CD45RO+記憶性T細(xì)胞、刺激過的初始和記憶性CD4+和CD8+T細(xì)胞、NK細(xì)胞、噬堿性粒細(xì)胞、噬酸性粒細(xì)胞、樹突狀細(xì)胞、肥大細(xì)胞、單核細(xì)胞和小膠質(zhì)細(xì)胞。除了對細(xì)胞遷移的影響外,RANTES還能活化多種細(xì)胞,包括T細(xì)胞、單核細(xì)胞、中性粒細(xì)胞、NK細(xì)胞、樹突狀細(xì)胞和星型膠質(zhì)細(xì)胞。
產(chǎn)品原理
本實驗采用雙抗體夾心ELISA法。抗人CCL5/Rantes單抗包被于酶標(biāo)板上,標(biāo)本和標(biāo)準(zhǔn)品中的CCL5/Rantes會與單抗結(jié)合形成免疫復(fù)合物,游離的成分被洗去;加入生物素化的抗人CCL5/Rantes抗體,生物素化抗人CCL5/Rantes抗體與酶標(biāo)板上結(jié)合的標(biāo)準(zhǔn)品或樣本中的CCL5/Rantes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CCL5/Rantes,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,CCL5/Rantes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CCL5/Rantes濃度。
產(chǎn)品性能
1. 靈敏度:最小可測人CCL5/Rantes達(dá)7pg/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 (60μl/支)
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1 支(60μl/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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4 瓶(25ml/瓶)
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2 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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3 張
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2 張
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10.說明書
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1 份
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1 份
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試驗所需自備儀器和試劑:
1. 酶標(biāo)儀(450nm檢測波長濾光片,570nm或630nm校正波長濾光片)。
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、坐標(biāo)紙。
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