التفاصيل البيبلوغرافية
| العنوان: |
A Robust Method for Packing High Resolution C18 RP-nano-HPLC Columns. |
| المؤلفون: |
Jami-Alahmadi Y; Department of Biological Chemistry, David Geffen School of Medicine at UCLA; yjami@ucla.edu., Pandey V; Department of Biological Chemistry, David Geffen School of Medicine at UCLA., Mayank AK; Department of Biological Chemistry, David Geffen School of Medicine at UCLA., Wohlschlegel JA; Department of Biological Chemistry, David Geffen School of Medicine at UCLA. |
| المصدر: |
Journal of visualized experiments : JoVE [J Vis Exp] 2021 May 14 (171). Date of Electronic Publication: 2021 May 14. |
| نوع المنشور: |
Journal Article; Research Support, N.I.H., Extramural; Video-Audio Media |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: |
Chromatography, High Pressure Liquid* , Chromatography, Reverse-Phase* , Peptides*/chemistry, Chromatography, Liquid ; Mass Spectrometry |
| مستخلص: |
The high complexity prevalent in biological samples requires chromatographic separations with high sensitivity and resolution to be effectively analyzed. Here we introduce a robust, reproducible and inexpensive protocol for preparation of a nano-flow reversed phase high performance liquid chromatography (RP-HPLC) columns for on-line separation of analytical peptides before introduction into and detection by a mass-spectrometer in traditional bottom-up proteomics workflows. Depending on the goal of the experiment and the chemical properties of the analytes being separated, optimal column parameters may differ in their internal or outer diameters, length, particle size, pore size, chemistry of stationary phase particles, and the presence or absence of an integrated electrospray emitter at the tip. An in-house column packing system not only enables the rapid fabrication of columns with the desired properties but also dramatically reduces the cost of the process. The optimized protocol for packing a C18 AQ (aqueous) fused silica column discussed here is compatible with a wide range of liquid chromatographic instruments for achieving effective separation of analytes. |
| المشرفين على المادة: |
0 (Peptides) |
| تواريخ الأحداث: |
Date Created: 20210531 Date Completed: 20210930 Latest Revision: 20210930 |
| رمز التحديث: |
20240628 |
| DOI: |
10.3791/62380 |
| PMID: |
34057454 |
| قاعدة البيانات: |
MEDLINE |
