Are the EasyPep MS Sample Prep Kits compatible with vacuum pumps/dry-down devices?
As per protocol, peptide samples need to be dried down prior to LC-MS analysis. This step can be performed using vacuum pumps/dry-down devices without any harm caused to the device by the components present in the elution buffer within the EasyPep MS Sample Prep Kits.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
After cell lysis, my mass spectrometry sample is very viscous and difficult to pipette. How do I reduce the sample viscosity?
High sample viscosity after lysis is due to release of DNA from the nucleus. Sonication or addition of a nuclease such as the Pierce Universal Nuclease (Cat. No. 88700, 88701, or 88702) can be used to degrade DNA and reduce sample viscosity.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
How do I process yeast samples with EasyPep Mini MS Sample prep Kit?
Add 0.5 mm diameter glass beads to the sample pelleted in Easy Pep Lysis Solution and proceed as recommended in the Easy Pep sample prep protocol. Vortex vigorously for few minutes, spin at 16,000 x g for 5 mins, and extract the supernatant for protein assay and further downstream sample preparation.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
How do I process bacterial samples with EasyPep MS Sample Prep kits?
Add 1-2 µL of 50 mg/mL Lysozyme Solution (Cat. No. 90082) to 200 µL of the EasyPep lysis buffer provided in the kit and proceed with the sample prep. Add 200 µL of the prepared lysis solution to 5 X 10E8 bacterial cells (E. coli cells) (OD600 equal to 1, corresponding to 1 X 10E8 CFUs). Lyse by pipetting the sample repeatedly and centrifuge at 16,000 x g for 5 mins. Alternatively, E. coli cells can be lysed in lysis buffer using bead beating or sonication instead of lysozyme. After lysis, dilute samples to 1 mg/mL using lysis buffer, proceed with the EasyPep protocol for reduction, alkylation, digestion, and clean up as described in the product manual.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
Do you offer a mass spec sample prep method for low abundant proteins in samples?
High Select Top14 Abundant Protein Depletion Mini Spin columns (Cat. Nos. A36369, A36370) can be used to deplete top 14 abundant plasma/serum proteins. The depletion of highly abundant proteins from the sample enables the detection and identification of low abundant proteins. The depleted samples can be processed with EasyPep MS sample prep kit to generate digested plasma/serum samples. The clean digested samples can be processed by nanoLC-MS/MS analysis for discovery experiments or nanoLC-PRM/MS analysis for targeted experiments.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.