ReadiLink™xtra Rapid AF647抗体标记试剂盒

货号1985存储条件 在零下15度以下保存, 避免光照
规格2 Labelings价格2412
Ex (nm)650Em (nm)671
分子量溶剂
产品详细介绍


简要概述

ReadiLink™xtra快速抗体标记试剂是AAT 研发,由百萤代理销售的AF647标记的抗体标记试剂盒。ReadiLink™xtra快速抗体标记试剂盒基本上只需要2个简单的混合步骤,而无需纯化。ReadiLink™试剂盒中使用的预活化AF647非常稳定,对抗体表现出良好的反应性和选择性。该试剂盒具有用于标记〜2x50 ug抗体的所有基本成分。试剂盒中提供的两个预活化AF647染料小瓶中的每一个均经过优化,可标记约50 µg抗体。ReadiLink™xtra AF647快速抗体标记试剂盒提供了一种方便而强大的方法,可以使用红色的AF647荧光团标记单克隆和多克隆抗体。百萤生物是AAT Bioquest的中国代理商,为您提供最优质的抗体标记试剂盒。 

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产品说明书

实验方案

工作溶液配制

蛋白质工作溶液(A)

为了标记50 µg蛋白质(假设目标蛋白质浓度为1 mg / mL),请将5 µL(反应总体积的10%)反应缓冲液(组分B)与50 µL目标蛋白质溶液混合。
注意:如果蛋白质浓度不同,请相应地调整蛋白质体积,以使〜50 µg蛋白质可用于标记反应。
注意:要标记100 µg蛋白质(假设目标蛋白质浓度为1 mg / mL),请将10 µL(反应总体积的10%)反应缓冲液(组分B)与100 µL目标蛋白质溶液混合。
注意:蛋白质应溶于1X磷酸盐缓冲盐水(PBS),pH 7.2-7.4;如果蛋白质溶解在甘氨酸缓冲液中,则必须针对1X PBS(pH 7.2-7.4)进行透析,或使用10 kDa的Amicon Ultra-0.5,Ultracel-10膜去除游离的胺或铵盐(例如硫酸铵和乙酸铵)被广泛用于蛋白质沉淀。
注意:为获得最佳标记效率,建议最终蛋白质浓度范围为1-2 mg / mL,结合效率明显降低,低于1 mg / mL。 

 

操作步骤

运行缀合反应
  1. 将蛋白质工作溶液(溶液A)添加到一个小瓶的标记染料(组分A)中,并通过将小瓶涡旋几秒钟将它们充分混合。
    注意:如果要标记100 µg的蛋白质,请使用两个小瓶(组分A),将100 µg的蛋白质分成2 x 50 µg的蛋白质,并使每个50 µg的蛋白质与一小瓶的标记染料反应。然后合并两个小瓶,用于下一步。
  2. 将缀合反应混合物在室温下放置30-60分钟。
    注意:如果需要,可以旋转或摇动缀合反应混合物更长的时间。 

 

停止缀合反应
  1. 将5 µL(对于50 µg蛋白质)或10 µL(对于100 µg蛋白质)添加到缀合反应混合物中,占TQ™染色猝灭缓冲液(组分C)总反应体积的10%;混合均匀。
  2. 在室温下孵育10分钟。标记的蛋白(抗体)可以使用了。 

 

蛋白质缀合物的储存

蛋白质缀合物应在载体蛋白(例如0.1%牛血清白蛋白)存在下以> 0.5 mg / mL的浓度存储。为了更长的存储时间,可以将蛋白质缀合物冻干或分成单份使用,并存储在≤–20°C下。

 

图示

图1. HeLa细胞中微管蛋白的免疫荧光染色。HeLa细胞用4%PFA固定,用0.1%Triton X-100透化并封闭。然后将细胞与小鼠抗微管蛋白抗体一起孵育,并用ReadiLink™xtra Rapid AF647抗体标记试剂盒(#1985)标记的山羊抗小鼠IgG染色。

  

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说明书
ReadiLink™xtra Rapid AF647抗体标记试剂盒.pdf