کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
4337188 | 1614661 | 2006 | 4 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Phospho-CREB and other phospho-proteins: Improved recovery from brain tissue Phospho-CREB and other phospho-proteins: Improved recovery from brain tissue](/preview/png/4337188.png)
During attempts to quantify levels of phosphorylated cAMP response element binding protein (CREB-P) in guinea pig brain stem auditory nuclei by Western blotting, we compared the decay of CREB-P levels when tissues were homogenized in traditional Lysis buffer containing detergents or in 50 mM Tris–HCl buffer containing 0.32 M sucrose. The decay of CREB-P levels was retarded considerably in the Tris–Sucrose medium as compared to the Lysis buffer. Similarly, the levels of two other phospho-proteins, extracellular regulated kinases (ERK1/2-P) and stress activated protein/Jun-N-terminal kinase (SAP/JNK-P), were better preserved by the Tris–Sucrose medium. These findings imply that the detergents typically present in the Lysis buffer may disrupt organelles and increase the exposure of soluble phospho-proteins to hydrolyzing enzymes. In contrast, such exposure was probably minimized in the Tris–Sucrose medium, which is thought to preserve organelle integrity.
Journal: Journal of Neuroscience Methods - Volume 150, Issue 2, 30 January 2006, Pages 238–241