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.
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I have a lot of plasma samples to prepare for mass spectrometry analysis. Is there a way to perform abundant protein depletion in a 96-well plate?
The bulk Top14 depletion resin is compatible with a 96-well filter plate such as the Agilent filter microplate (Cat. No. 200957-100). Add 600 µL of resin slurry (50%) to each of the wells in the 96-well filter plate, add 10-20 µL of sample, and incubate with gentle shaking. Depleted samples can be collected in a new 96-well polypropylene plate by centrifugation for 2 mins at 1,000 x g or by using a vacuum manifold.
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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.
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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.
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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.
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