| Measurement of ATP Hydrolytic Activity of Plasma Membrane H+-ATPase from Arabidopsis thaliana Leaves
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Author:
Date:
2016-12-05
[Abstract] Plant plasma membrane H+-ATPase, which is a P-type ATPase, couples ATP hydrolysis to H+ extrusion and thereby generates an electrochemical gradient across the plasma membrane. The proton gradient is necessary for secondary transport, cell elongation, and membrane potential maintenance. Here we describe a protocol for measurement of the ATP hydrolytic activity of the plasma membrane H+-ATPase from Arabidopsis thaliana leaves.
[摘要] 作为P型ATP酶的植物质膜H sup + -ATPase将ATP水解耦合到H + +/- 挤出，从而在质膜上产生电化学梯度。质子梯度对于二次转运，细胞伸长和膜电位维持是必需的。这里我们描述用于测量来自拟南芥叶片的质膜H + -ATPase的ATP水解活性的方案。
关键词： strong> 拟南芥，ATP水解活性，原钒酸盐，P-型ATP酶，血浆膜H + -ATPase
质膜H + -ATPase活性的测定对于阐明其功能和调节机制是重要的。然而，有时难以确定质膜H sup + -ATP酶的ATP水解活性，因为植物细胞含有许多ATP水解酶。该协议是基于Uemura和Yoshida（1986）和Kinoshita等人的出版物开发的。 （1995）。我们使用KNO 3作为V型ATP酶的抑制剂，钼酸铵作为酸性磷酸酶的抑制剂，寡霉素作为F型ATP酶的抑制剂，NaF作为磷酸酶的抑制剂（Shimazaki和Kondo ，1987; Kinoshita等人，1995）。原钒酸盐抑制P型ATP酶，并且因此可以通过评估来自ATP水解的钒酸盐敏感性P 1释放来用于测量质膜H sup + -ATP酶的活性。释放的P 1与钼酸盐反应形成蓝色络合物，然后可以通过测量在750nm的吸收来量化。
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| ELISA Detection of Endogenous Serum Albumin in the Mouse Brain: A Measure of Extravasation Following Brain Injury
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Author:
Date:
2015-05-05
[Abstract] After stroke and brain contusion, serum proteins extravasate into nerve tissue through disrupted blood-brain barrier (BBB). Because extravasations of serum proteins result in vasogenic brain edema, serum albumin level in the brain is an indicator of BBB disruption and brain edema after brain insults. In this protocol, extravasation of endogenous albumin is measured in the damaged mouse brain, which would be valuable in the evaluation of vasogenic brain edema formation (Michinaga et al., 2014).
[摘要] 在中风和脑挫伤后，血清蛋白通过破裂的血脑屏障（BBB）外渗到神经组织中。 因为血清蛋白的外渗导致血管性脑水肿，脑中的血清白蛋白水平是脑损伤后BBB破坏和脑水肿的指标。 在该方案中，在损伤的小鼠脑中测量内源性白蛋白的外渗，这在评价血管性脑水肿形成中是有价值的（Michinaga等人，2014）。
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