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验证六种商业抗体检测异源和内源性 TRPM8 离子通道表达。

Validation of Six Commercial Antibodies for the Detection of Heterologous and Endogenous TRPM8 Ion Channel Expression.

机构信息

Instituto de Neurociencias (IN), CSIC-UMH, 03550 San Juan de Alicante, Spain.

出版信息

Int J Mol Sci. 2022 Dec 18;23(24):16164. doi: 10.3390/ijms232416164.

Abstract

TRPM8 is a non-selective cation channel expressed in primary sensory neurons and other tissues, including the prostate and urothelium. Its participation in different physiological and pathological processes such as thermoregulation, pain, itch, inflammation and cancer has been widely described, making it a promising target for therapeutic approaches. The detection and quantification of TRPM8 seems crucial for advancing the knowledge of the mechanisms underlying its role in these pathophysiological conditions. Antibody-based techniques are commonly used for protein detection and quantification, although their performance with many ion channels, including TRPM8, is suboptimal. Thus, the search for reliable antibodies is of utmost importance. In this study, we characterized the performance of six TRPM8 commercial antibodies in three immunodetection techniques: Western blot, immunocytochemistry and immunohistochemistry. Different outcomes were obtained for the tested antibodies; two of them proved to be successful in detecting TRPM8 in the three approaches while, in the conditions tested, the other four were acceptable only for specific techniques. Considering our results, we offer some insight into the usefulness of these antibodies for the detection of TRPM8 depending on the methodology of choice.

摘要

TRPM8 是一种非选择性阳离子通道,表达于初级感觉神经元和其他组织中,包括前列腺和尿路上皮。其参与体温调节、疼痛、瘙痒、炎症和癌症等不同生理和病理过程的作用已被广泛描述,使其成为治疗方法的一个有前途的靶点。TRPM8 的检测和定量对于深入了解其在这些生理病理条件下的作用机制至关重要。基于抗体的技术常用于蛋白质的检测和定量,但它们在许多离子通道(包括 TRPM8)上的性能并不理想。因此,寻找可靠的抗体至关重要。在这项研究中,我们在三种免疫检测技术:Western blot、免疫细胞化学和免疫组织化学中,对六种 TRPM8 商业抗体的性能进行了表征。对测试的抗体得到了不同的结果;其中两种抗体在三种方法中都能成功检测到 TRPM8,而在测试的条件下,另外四种抗体仅在特定的技术中可用。考虑到我们的结果,我们根据所选方法,为这些抗体在检测 TRPM8 方面的有用性提供了一些见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a803/9784522/1ddc0e96d12c/ijms-23-16164-g001.jpg

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