مورد إلكتروني
Detection of tyrosine phosphorylated proteins by combination of immunoaffinity enrichment, two-dimensional difference gel electrophoresis and fluorescent Western blotting
| العنوان: | Detection of tyrosine phosphorylated proteins by combination of immunoaffinity enrichment, two-dimensional difference gel electrophoresis and fluorescent Western blotting |
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| بيانات النشر: | Institutionen för genetik och patologi 2010 |
| تفاصيل مُضافة: | Lind, Sara Bergström Hagner-McWhirter, Åsa Elfineh, Lioudmila Molin, Magnus Jorsback, Anneli Öhman, Johan Pettersson, Ulf |
| نوع الوثيقة: | Electronic Resource |
| مستخلص: | We describe fluorescence-based 2-D gel electrophoresis methods for visualization of low abundant cancer relevant tyrosine phosphorylated (pTyr) proteins The methods investigated were fluorescent Western blotting and two-dimensional difference gel electrophoresis (2-D DIGE) for detection of non-enriched and immunoaffinity enriched pTyr protein patterns The same anti-phosphotyrosine specific antibody 4G10 was used for both approaches The results from fluorescent Western blotting of total proteins and from enriched CyDye DIGE pre-labeled pTyr proteins showed similar down regulation of phosphorylation upon treating of cells from a cancer model system (K562 chronic myeloid leukemia cells) with imatinib This treatment introduced a known perturbation of phosphorylation that enabled testing of these new approaches to analyze variations in tyrosine phosphorylation levels Enrichment of pTyr proteins was found highly advantageous for the outcome Out of a simplified 2 D DIGE experiment of immunoaffinity enriched control and treated pTyr proteins differential analysis as well as protein identification by mass spectrometry (MS) was possible. |
| مصطلحات الفهرس: | Fluorescence, Fluorescent Western blotting, Proteomics, Two-dimensional difference gel electrophoresis, Tyrosine phosphorylation, Medical and Health Sciences, Medicin och hälsovetenskap, Article in journal, info:eu-repo/semantics/article, text |
| DOI: | 10.1016.j.bbrc.2010.09.104 |
| URL: | Biochemical and Biophysical Research Communications - BBRC, 0006-291X, 2010, 401:4, s. 581-585 |
| الإتاحة: | Open access content. Open access content info:eu-repo/semantics/restrictedAccess |
| ملاحظة: | English |
| أرقام أخرى: | UPE oai:DiVA.org:uu-135372 doi:10.1016/j.bbrc.2010.09.104 ISI:000283972400016 1235104868 |
| المصدر المساهم: | UPPSALA UNIV LIBR From OAIster®, provided by the OCLC Cooperative. |
| رقم الأكسشن: | edsoai.on1235104868 |
| قاعدة البيانات: | OAIster |
| DOI: | 10.1016.j.bbrc.2010.09.104 |
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