哮喘小鼠肺中常用管家蛋白的表达发生改变

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Abigail R. Patterson , Alexandra P. Crawford , Amanda S. Hatcher , Dovenia S. Ponnoth , Shinichi Asano
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引用次数: 0

摘要

组织样品中的管家蛋白表达通常用于标准化免疫印迹数据。然而,在没有验证的病理条件下使用常见的负载控制蛋白可能是有问题的。在这里,我们描述了哮喘肺样本中常用的负载控制蛋白表达的变化,并提出了一种更简单、更可靠的western blot分析负载控制。来自对照组和哮喘小鼠组的肺样本用于评估三种常用的家政蛋白(β -肌动蛋白、α -微管蛋白和甘油醛-3-磷酸脱氢酶)的表达水平。还获得Ponceau S染色数据以评估蛋白质负载的一致性。管家蛋白在哮喘组中的表达差异显著,与对照组相比,变化幅度为26%至278% (p <;0.05)。另一方面,各组间Ponceau S染色无显著差异。各组肺标本蛋白提取量无差异,但支气管肺泡灌洗白细胞群有显著差异(p <;0.05)。我们的数据表明,哮喘组的肺常驻免疫细胞发生了显著改变,而常用的负荷控制蛋白的蛋白表达对于哮喘肺的western blot分析是不可靠的。我们推荐Ponceau S染色作为哮喘肺样本的更可靠的负荷对照,以使western blot数据正常化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Expressions of commonly used housekeeping proteins are altered in asthmatic mouse lungs
Housekeeping protein expression in tissue samples is often used to normalize immunoblotting data. However, utilizing common loading control proteins in pathological conditions without validation can be problematic. Here, we describe the alteration of commonly used loading control protein expressions in asthmatic lung samples and present a simpler and more reliable loading control for western blot analysis. Lung samples from control and asthmatic mice groups were used to assess the expression levels of three commonly used housekeeping proteins (beta-actin, alpha-tubulin, and glyceraldehyde-3-phosphate dehydrogenase). Ponceau S staining data was also obtained to assess the consistency of protein loading. Housekeeping protein expressions varied dramatically in asthmatic groups, ranging from 26 to 278 % compared to the control group (p < 0.05). On the other hand, no significant differences in Ponceau S staining were observed among the groups. There were no differences in protein extraction from lung samples among the groups, but significant differences were observed in bronchoalveolar lavage leukocyte populations (p < 0.05). Our data suggest that lung resident immune cells were dramatically altered in asthma groups, and protein expression of commonly used loading control proteins are unreliable for western blot analysis of asthmatic lungs. We recommend Ponceau S staining as a more reliable loading control for asthmatic lung samples to normalize western blot data.
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来源期刊
Biochemistry and Biophysics Reports
Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics
CiteScore
4.60
自引率
0.00%
发文量
191
审稿时长
59 days
期刊介绍: Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.
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