Pierce Graphite Spin Columns 石墨旋转柱
货号：88302
规格：30 columns
价格：1419
产品类型：蛋白表达研究
品牌：Thermo Fisher
| Thermo Scientific Pierce Graphite Spin Columns improve phosphopeptide analysis by efficiently binding hydrophilic peptides and efficiently removing urea, salts and other contaminants before mass spectrometry analysis.Pierce Graphite Spin Columns enable fast and efficient capture, concentration, desalting and elution of hydrophilic peptides in less than 10 minutes. These columns are ideal for improving mass spectrometric (MS) analyses of samples from protein digests), strong-cation exchange fractions, and enriched phosphopeptides eluted from titanium dioxide and immobilized metal affinity chromatography (IMAC) columns and tips.The Pierce Graphite Spin Columns improve phosphopeptide analysis by efficiently binding hydrophilic peptides and efficiently removing urea, salts and other contaminants before MS analysis. The C18 resins and C18 tips that are commonly used to desalt peptides are excellent for use with hydrophobic peptides but do not efficiently capture hydrophilic peptides, like phosphopeptides, resulting in enrichment of only hydrophobic fragments. The Pierce Graphite Spin Columns address this issue and are ideal for matrix-assisted laser desorption ionization (MALDI) or nanoelectrospray ionization techniques. |
| 特点： |
| - Convenient spin format for parallel processing of multiple samples- High-binding capacity with excellent recovery of up to 100 µg of hydrophilic peptides per column- Porous graphite resin for cleaning up phosphopeptide samples before MS analysis |
| 数据： |
Graphite clean-up enables phosphopeptide identificationU2OS human osteosarcoma cells synchronized at the G2/M boundary with nocodazole (200ng/mL, 36 hours) were lysed with 6M guanidine•HCl. After enzymatic protein digestion (100µg), phosphopeptides were enriched with IMAC and desalted with Pierce Graphite Spin Columns or C18 tips before LC-MS/MS analysis on a Thermo Scientific Orbitrap XL Mass Spectrometer. Two representative spectra are shown for two phosphopeptides not observed after C18 cleanup. Panel A: A novel doubly-phosphorylated peptide was identified within the putative ATP binding site of cyclin dependent kinase cdc2 (CDK1). This phosphopeptide is not present in Phospho.ELM version 8.2 database. Panel B: Dual specificity mitogen-activated protein kinase kinase 2 (MP2K2) phosphopeptide. Improved recovery of representative hydrophilic phosphopeptides using graphite spin columnsStable isotope-labeled A3 and B9 peptides (10pmol) were acidified with 1% trifluoroacetic acid, processed according to instructions for C18 tips or the Pierce Graphite Spin Columns and eluted with 50% acetonitrile with 0.1% formic acid. The corresponding heavy isotope labeled peptides (5pmol) were spiked in the eluate, dried and resuspended in 0.1% formic acid. Samples were analyzed by targeted LC-MS/MS with the Thermo Scientific Orbitrap XL Mass Spectrometer to quantitate percent recovery.Peptides:A3 = RPRAApTFPFR¹B9 = RTPKDpSPGIPPFR¹¹Position of heavy isotope labeled amino acid used for absolute MS quantitation. |
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