二抗是在其它宿主体内制备的能与一抗或一抗片段结合的抗体,根据实验需求,二抗上通常连有酶或荧光素等标签。由于二抗所具备的优点使得其在免疫学实验中得以广泛应用,如WB/ELISA/IHC/IP等。二抗针对某一特定物种的所有抗体均具有特异性,因而使用特定标记的二抗可以免去对每一个一抗进行标记,大大节省了时间和费用;此外,一个一抗分子可以同时结合几个二抗分子,从而使信号大大增强,提高了实验灵敏度。
二抗,广义上是指专门和一抗进行特异性反应和结合的抗体。检测任何目的靶蛋白都有不止一种抗体可供选择,同样的,在选择二抗的时候要综合考虑一抗的类型及后继检测方案的要求,一般来说,选择合适的二抗需要从以下几个方面考虑:
1.一抗的种属来源
根据一抗的物种来源选择相应的抗该物种的二抗。例如:一抗是小鼠源的单克隆抗体,二抗则选抗小鼠的二抗(山羊抗小鼠或兔抗小鼠等)。
2.一抗的类别亚型
二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白,因此相应的二抗就是抗IgG抗体。单克隆抗体的类别及亚型通常会在产品说明书中有描述,如果一抗是小鼠IgM,那么相应的二抗就应当是抗IgM。如果单克隆一抗是小鼠IgG的某一亚型(IgG1、IgG2a、IgG2b、IgG3),那么几乎所有的抗小鼠IgG都可以与之结合,或者你也可以选择专门针对这一亚型的二抗。此种抗体在双标记实验中尤其合适。
3.二抗的偶联标记
一般来讲,偶联到二抗上的探针主要有酶(辣根过氧化酶HRP)、荧光基团、生物素、金颗粒等。选用哪种探针的二抗主要取决于具体的实验。WB、ELISA和IHC常用的二抗通常是酶标二抗(部分荧光WB需选用远红外波长荧光蛋白标记的荧光二抗),而免疫荧光、流式等实验中通常使用荧光基团标记的二抗,而金颗粒标记的二抗则更多的应用于免疫电镜中。 还有一些需要更大程度地放大信号的免疫实验中,二抗则可以选择biotin/avidin体系的二抗或无偶联标记的二抗(再用三抗放大)。
国产优质品牌AG8亚国服务为您提供全种类的高品质二抗供您选择,高品质的抗体可以使您的实验更省时省力的取得真实的结果,优质国产品牌AG8亚国服务,满足您的对高品质抗体的需求。具体产品可点击:
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AG8亚国服务特色产品线(全部现货):
二抗、标签抗体、对照抗体;WB相关:ECL发光液、预染marker、预制胶;IHC相关:二抗试剂盒、组化笔;IP/CoIP试剂盒;细胞因子、信号通路调节剂;血清、BSA、蛋白酶K、CTB、TTX、CEE;凋亡试剂盒、呼吸爆发试剂盒;动植物提取物,定制服务(抗体、多肽)...
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