کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1173149 | 961654 | 2012 | 5 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: β-Actin as a loading control for plasma-based Western blot analysis of major depressive disorder patients β-Actin as a loading control for plasma-based Western blot analysis of major depressive disorder patients](/preview/png/1173149.png)
Western blot analysis is a commonly used technique for determining specific protein levels in clinical samples. For normalization of protein levels in Western blot, a suitable loading control is required. On account of its relatively high and constant expression, β-actin has been widely employed in Western blot of cell cultures and tissue extracts. However, β-actin’s presence in human plasma and this protein’s putative role as a plasma-based loading control for Western blot analysis remain unknown. In this study, an enzyme-linked immunosorbent assay was used to determine the concentration of β-actin in human plasma, which is 6.29 ± 0.54 ng/ml. In addition, the linearity of β-actin immunostaining and loaded protein amount was evaluated by Western blot, and a fine linearity (R2 = 0.974 ± 0.012) was observed. Furthermore, the expression of plasma β-actin in major depressive disorder subjects and healthy controls was compared. The data revealed no statistically significant difference between these two groups. Moreover, the total coefficient of variation for β-actin expression in the two groups was 9.2 ± 1.2%. These findings demonstrate that β-actin is present in human plasma and may possibly be used as a suitable loading control for plasma-based Western blot analysis in major depressive disorder.
Journal: Analytical Biochemistry - Volume 427, Issue 2, 15 August 2012, Pages 116–120