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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What is the composition of the Lysis Solution in the EasyPep MS Sample Prep Kits (Cat. Nos. A4006, A45733, A5734, A45735)?
The composition of the Lysis Solution in the EasyPep MS Sample Prep Kits (Cat. Nos. A4006, A45733, A5734, A45735) is proprietary. The kit includes pre-formulated buffers, MS-grade enzyme mix, and peptide columns to prepare high-quality detergent-free peptide samples for direct LC-MS analysis or further sample processing such as isobaric tag (e.g., TMT or TMT pro reagent) labeling, phosphopeptide enrichment, or high-pH reversed-phase fractionation.
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Can I prepare my samples in Laemmli sample buffer for use with EasyPep MS Sample Prep Kits (Cat. Nos. A40006, A45733, A45734)?
We have not validated our EasyPep MS Sample Prep Kits (Cat. Nos. A40006, A45733, A45734) for use with Laemmli sample buffer. The lysis buffer provided with the kit has been optimized to be compatible with digestion and clean-up steps. We do not recommend using other buffers as they may contain detergents or other components that may interfere with reduction, alkylation, digestion, or peptide clean-up. The high concentration of SDS in Laemmli sample buffer could potentially disrupt the activity of the universal nuclease.
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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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