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DU-730 Spectrophotometer

NanoDrop

Company: Beckman Coulter
Catalog#: DU 730
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Extraction and Measurement of Abscisic Acid in a Unicellular Red Alga Cyanidioschyzon merolae
Author:
Date:
2016-12-05
[Abstract]  Abscisic acid (ABA) has been known as a phytohormone of land plants, which is synthesized in response to abiotic stresses and induces various physiological responses, but is also found from eukaryotic algae. Recently, we reported that a unicellular red alga Cyanidioschyzon merolae produced ABA, which prevented cell growth and enhanced salt stress tolerance (Kobayashi et al., 2016). This report describes the detailed method for the extraction and quantification of ABA in the model red alga C. merolae. [摘要]  脱落酸(ABA)已知为陆生植物的植物激素,其响应于非生物胁迫合成并诱导各种生理反应,但也可从真核藻类中发现。最近,我们报道了单细胞红藻(Cyanidioschyzon merolae)产生ABA,其阻止细胞生长和增强盐胁迫耐受性(Kobayashi等人,2016)。该报告描述了在红海藻模型中提取和定量ABA的详细方法。 。
关键字:脱落酸,藻类, Cyanidioschyzon merolae ,HPLC,植物激素

] 植物激素ABA已在发散光合真核生物中发现,但单细胞藻类的功能仍不清楚。在最近的研究中,我们显示单细胞红藻。 melorae 通过本方案累积ABA以应答盐胁迫。这是用于从C中提取和定量ABA的第一已公布方案的细节。 merolae 。此协议针对 C进行了优化。基于陆地植物协议的。

Measurement of ATP Hydrolytic Activity of Plasma Membrane H+-ATPase from Arabidopsis thaliana Leaves
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。

DNA Slot Blot Repair Assay
Author:
Date:
2015-04-20
[Abstract]  Ultraviolet (UV) irradiation induces helix distorting photolesions such as cyclobutane pyrimidine dimers (CPD) and pyrimidine-pyrimidone (6-4) photoproducts (6-4PP) which threaten genomic integrity if unrepaired. In mammals, nucleotide excision repair (NER) is the only pathway that removes UV-induced DNA damages. Here we describe DNA slot blot repair assay for quantitative detection of NER activity using DNA damage specific antibodies such as anti-CPD and anti-6-4PP. Briefly, genomic DNA irradiated with UV was isolated from cells, and the genomic DNA was vacuum-transferred to a nitrocellulose membrane using a Bio-Dot SF microfiltration apparatus (Bio-Rad). A monoclonal antibody that recognizes CPD or 6-4PP was applied to detect the remaining amount of photolesions in the genomic DNA. For ... [摘要]  紫外线(UV)照射诱导螺旋扭曲光致损伤,例如环丁烷嘧啶二聚体(CPD)和嘧啶 - 嘧啶酮(6-4)光产物(6-4PP),如果未修复则威胁基因组完整性。 在哺乳动物中,核苷酸切除修复(NER)是去除UV诱导的DNA损伤的唯一途径。 在这里我们描述DNA狭缝印迹修复测定NER活性使用DNA损伤特异性抗体如抗CPD和抗6-4PP的定量检测。 简言之,从细胞中分离用UV照射的基因组DNA,使用Bio-Dot SF微量过滤装置(Bio-Rad)将基因组DNA真空转移到硝酸纤维素膜上。 应用识别CPD或6-4PP的单克隆抗体来检测基因组DNA中残留的光损伤量。 对于均匀负载的上样控制,可以通过SYBR金染色进一步分析DNA在膜上。

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