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Phosphatase substrate

磷酸酶底物

公司名称: Sigma-Aldrich
产品编号: S0942
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Cyclic Nucleotide (cAMP and cGMP) Assays and Capture ELISA for Quantitative Analysis of Plasmodium falciparum Blood-stage Egress
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2014-03-05
[Abstract]  Upon rupture of Plasmodium falciparum (P. falciparum) schizonts in vitro (an event known as egress), merozoites are released into the culture medium. The merozoites invade fresh red blood cells, a process that involves shedding of a microneme protein called apical membrane antigen-1 (AMA1) from the merozoite surface. This shedding, which takes place even in the absence of invasion, is therefore a surrogate marker for the degree of egress taking place in a culture, and can be measured using a specific capture ELISA to quantify AMA1 levels in culture supernatants ... [摘要]  在体外破裂镰状疟原虫(恶性疟原虫)裂殖体(一种称为外出的事件)时,裂殖子释放到培养基中。裂殖子侵入新鲜的红细胞,该过程涉及从裂殖子表面脱落称为顶膜抗原-1(AMA1)的微米蛋白蛋白质。因此,即使在不存在侵袭的情况下发生的脱落也是培养物中出现的出口程度的替代标记,并且可以使用特异性捕获ELISA来测量,以定量培养物上清液中的AMA1水平(Collins, et al。,2013)。该测定使用特异于AMA1的单克隆抗体(称为4G2dc1)(Kocken等人,1998; Collins等人,2009),以捕获和固定来自培养物的蛋白质然后使用特异性兔多克隆抗血清来检测固定的抗原。最终使用磷酸酶缀合的山羊抗兔抗体来定量第二抗体的结合。出口绝对依赖于寄生虫cGMP依赖性蛋白激酶PKG的活性,因此受细胞内cGMP水平的影响(Collins等人,2013)。这由鸟苷酸环化酶和磷酸二酯酶之间的相互作用调节。后者酶也可以降解cAMP,因此它也可以是信息性的测量细胞内cAMP水平

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