Read e-book online Quantitative Proteomics by Mass Spectrometry (Methods in PDF

By Salvatore, Ed. Sechi

ISBN-10: 1588295710

ISBN-13: 9781588295712

ISBN-10: 1597452556

ISBN-13: 9781597452557

Show description

Read Online or Download Quantitative Proteomics by Mass Spectrometry (Methods in Molecular Biology Vol 359) PDF

Best biology books

Download e-book for iPad: Apoptosis Methods and Protocols by Demetrius Matassov, Terri Kagan, Julie Leblanc (auth.), Hugh

A suite of state-of-the-art options for detecting and quantifying apoptosis, knowing its biochemistry, and for opting for the genes and proteins that keep watch over and hold it out. defined in step by step element, those simply reproducible equipment variety from circulate cytometry and immunohistochemical methods to kinase job assays, yeast two-hybrid screening, and the cloning of novel genes by means of differential expression.

Download e-book for iPad: Cellular and Molecular Biology of Metals by Rudolfs K. Zalups, D. James Koropatnick

With bankruptcy contributions from greater than 30 steel biology specialists, mobile and Molecular Biology of Metals explains the function of key divalent steel ions desirous about the molecular and mobile biology of assorted aim mobilephone populations. even though it basically specializes in homeostatic metals, equivalent to nickel, zinc, and chromium, the textual content additionally discusses a number of environmentally pertinent, poisonous divalent cations, together with mercury, cadmium, and arsenic.

Read e-book online The Cytoskeleton: An Introductory Survey PDF

The time period cytoskeleton has turn into firmly validated in state-of-the-art medical vocabulary. certainly, it truly is tough to think that merely ten years in the past, it used to be nearly non-existent. because then, the trendy box of study at the structural association of the cytoplasm has changed into essentially the most effective and swiftly increasing study parts in mobilephone Biology at the present time.

Extra info for Quantitative Proteomics by Mass Spectrometry (Methods in Molecular Biology Vol 359)

Sample text

3. Stable Isotope Labeling With ICAT Reagents 1. Eluted proteins from the p18 purifications (each containing approx 3 µg of total protein) are concentrated to 25 µL each in Nanosep 3 K centrifugal devices (see Note 6). 2. 3 and 1 mM EDTA. Keep an aliquot for analysis by silver-stained SDS-PAGE. 3. 3% and heating to 37°C (see Note 11). 4. The samples are reduced by addition of TBP to 5 mM and incubation at 37°C for 30 min. 05% SDS and 6 M urea. 5. One tube of heavy and one tube of light ICAT reagent are briefly spun in a microfuge to pellet the reagent.

4. Harvest of Cells, Lysis, and Estimation of Total Protein 1. Cells can be harvested using any standard protocol in tissue culture. In this example using HeLa cells, aspirate the medium from the cells and rinse the attached cells in warm phosphate buffered saline to remove remaining serum proteins. This is done twice to reduce the amount of serum albumin (a major component of serum) detectable in subsequent MS analyses. Scrape the cells in a small volume of lysis buffer (750 µL for a 14-cm dish), vortex frequently to ensure cell lysis, and obtain a small aliquot of each cell lysate for a Bradford assay to estimate protein concentration.

Monomeric avidin cartridge (Applied Biosystems) (see Note 3). 3. 4% trifluoroacetic acid (TFA). Prepare fresh. 4. Avidin wash buffer: 50 mM ammonium bicarbonate and 20% methanol. Prepare fresh. 5. Glass collection vials (Waters, Milford, MA; cat. no. WAT025054). Fig. 1 (continued from opposite page) the samples are prepared identically. Affinitypurified proteins from specific and control purifications are reduced, labeled with either the isotopically heavy or normal version of the ICAT reagent, and combined.

Download PDF sample

Quantitative Proteomics by Mass Spectrometry (Methods in Molecular Biology Vol 359) by Salvatore, Ed. Sechi


by Jeff
4.4

Rated 4.49 of 5 – based on 24 votes
This entry was posted in Biology.