The iDimerize Reverse Dimerization System brings the disruption of protein complexes under real-time, small-molecule control. A protein of interest is fused to the DmrD binding domain, and the fusion protein molecules aggregate unless the D/D Solubilizer ligand is present. Plasmid and lentiviral (Lenti-X) vector formats are available.
The iDimerize Reverse Dimerization System brings the disruption of protein complexes under real-time, small-molecule control. A protein of interest is fused to the DmrD binding domain, and the fusion protein molecules aggregate unless the D/D Solubilizer ligand is present. Plasmid and lentiviral (Lenti-X) vector formats are available.
Reverse dimerization: Disrupting protein-protein interactions
The iDimerize Reverse Dimerization System is a “reverse dimerization” system—aggregation is the resting state, and the D/D Solubilizer breaks up protein-protein interactions. Therefore, the iDimerize Reverse Dimerization System complements inducible dimerization, and can be used in analogous ways to create inducible alleles. In principle, most processes that can be brought under dimerizer control can also be controlled in the reverse manner using this kit to turn off a process that is activated by oligomerization.
Inducible secretion
The ability to create large protein aggregates has unique applications. For example, adding a secretory signal sequence to fusion proteins allows them to be reversibly stored as aggregates in the endoplasmic reticulum. The ligand can then be added to induce a rapid pulse of protein secretion from the cells. This method has been used to induce rapid, transient, and tightly regulated secretion of human growth hormone (hGH) and insulin (Rivera et al. 2000).
Protein aggregates can also be used in protein trafficking research. For example, this approach has been used to discover the existence of “mega-vesicles” transporting cargo across the Golgi stack (Volchuk et al. 2000).
D/D Solubilizer ligand
The D/D Solubilizer is a synthetic, cell-permeable ligand that can be used to disrupt dimerization of fusion proteins containing the DmrD domain. The D/D Solubilizer has been tested in vitro and in mice. It is nontoxic. We suggest testing various D/D Solubilizer concentrations within the recommended range (10–500 nM) for different lengths of time (30 minutes to 12+ hours) in order to obtain a complete dose-response profile.
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1.单克隆抗体的优点:
(1)杂交瘤可以在体外“永久”地存活并传代,只要不发生细胞株的基因突变,就可以不断的生产高特异性、高均一性的抗体.
(2)可以用相对不纯的抗原,获得大量高度特异的、均一的抗体.
(3)由于可能得到“无限量”的均一性抗体,所以适用于以标记抗体为特点的免疫学分析方法,如IRMA和ELISA等.
(4)由于单克隆抗体的高特异性和单一生物学功能,可用于体内的放射免疫显像和免疫导向治疗.
2.单克隆抗体的局限性:
(1)单克隆抗体固定的亲和性和局限的生物活性限制了它的应用范围.由于单克隆抗体不能进行沉淀和凝集反应,所以很多检测方法不能用单克隆抗体完成.
(2)单克隆抗体的反应强度不如多克隆抗体.
(3)制备技术复杂,而且费时费工,所以单克隆抗体的价格也较高.
抗原有两个基本特性,即抗原性和免疫原性。有抗原性的物质不一定有免疫原性,所以由此引出半抗原和完全全抗原,半抗原必须经过经过一定的改造(偶联蛋白载体BSA,OVA或者HSA等大分子物质)方能成为完全。一般而言完全抗原分子量越大(大于10KDa),结构越复杂引起免疫反应的能力也就越强。
抗体就是能与特异性抗原结合的免疫球蛋白,抗体一般分为多克隆抗体和单克隆抗体,多克隆抗体能与抗原的多个表位结合。本篇主要讲述兔来源的多克隆抗体的生产步骤
多抗一般制备流程:完全抗原的准备→兔子的免疫→ 效价检测和终放→抗体亲和纯化→抗体的浓缩和保存。
又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.
一般来说多克隆的阳性率高一些,但出现假阳性的比例也高一些。
其次,察看次目的蛋白的存在形式,有没有多聚体形式及变构形式;
最后,查看多家抗体公司的DATA,看看别人的WB做出来的条带的位置。
根据你说的,特异识别多个组织中的同样大小的条带,我觉得很可能就是你的目的蛋白。