1
Agilent Nanospray and HPLC-Chip/MS Protein Identification Solutions
Sample Preparation Guide
Agilent Technologies
1
Overview of Sample Preparation Steps
with the Protein ID Solutions
Optional removal of high-abundance proteins
3
Concentration of low-abundance proteins
3
Protein fractionation
3
Digestion
4
Cleanup
4
Figure 1
gives an overview of the sample preparation portion of the Agilent
Nanospray and HPLC-Chip/MS Protein Identification Solutions. The
applicable chapters are indicated adjacent to the workflow.
The solution starts with an optional immunodepletion column or spin
cartridge to remove high-abundance proteins. This is followed by protein
concentration with Agilent Spin Concentrators for Proteins. The sample is
then fractionated by either one-dimensional (1D) or two-dimensional (2D) gel
electrophoresis. The proteins are optionally reduced and alkylated, and then
are digested with the Agilent Protein In-Gel Tryptic Digestion Kit, or via
in-solution digestion. Then salts are removed with the Agilent Peptide Cleanup
C18 Spin Tubes or Cleanup C18 Pipette Tips. This Sample Preparation Guide
documents the entire set of steps.
After this, the sample proceeds to LC separation and MS analysis (see the
LC/MS Application Guide).
WA R N I N G
Before you begin, be sure you understand all safety considerations and have read all
applicable material data safety sheets.
Summary :
1 Agilent Nanospray and HPLC-Chip/MS Protein Identification Solutions Sample Preparation Guide Agilent Technologies 1 Overview of Sample Preparation Steps with the Protein ID Solutions Optional removal of high-abundance proteins 3 Concentration of low-abundance proteins 3 Protein fractionation 3 Digestion 4 Cleanup 4 Figure 1 gives an overview of the sample preparation portion of the Agilent Nanospray and HPLC-Chip/MS Protein Identification Solutions.
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