کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6270375 | 1295205 | 2008 | 5 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
The use of total protein stains as loading controls: An alternative to high-abundance single-protein controls in semi-quantitative immunoblotting
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علم عصب شناسی
علوم اعصاب (عمومی)
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![عکس صفحه اول مقاله: The use of total protein stains as loading controls: An alternative to high-abundance single-protein controls in semi-quantitative immunoblotting The use of total protein stains as loading controls: An alternative to high-abundance single-protein controls in semi-quantitative immunoblotting](/preview/png/6270375.png)
چکیده انگلیسی
Western blots are used to estimate the relative concentrations of proteins of interest based on staining by specific antibodies. Quantitative measurements are often subject to error due to overloading of the loading control and over-reliance on normalization. We have found that at the protein concentrations normally used to quantify most low-abundance proteins of interest, frequently used single-protein loading controls, such as glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and β-actin, do not accurately reflect differences in protein concentration. Two total protein stains, SYPRO® Ruby and Amido Black, were compared and found to be acceptable alternatives to single-protein controls. Although we cannot prove that high-abundance loading controls are inaccurate under all possible conditions, we conclude that the burden of proof should lie with the researcher to demonstrate that their loading control is reflective of quantitative differences in protein concentration.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Neuroscience Methods - Volume 172, Issue 2, 30 July 2008, Pages 250-254
Journal: Journal of Neuroscience Methods - Volume 172, Issue 2, 30 July 2008, Pages 250-254
نویسندگان
Georgina M. Aldridge, David M. Podrebarac, William T. Greenough, Ivan Jeanne Weiler,