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The following suggested protocol has been optimized using maximum and minimum protein concentrations of 0.5 mg/mL and 0.01 mg/mL respectively and is provided to demonstrate the potential uses of the ProTrap XG.
Our team works with customers to optimize protocols for your workflow, sample type, and research goals. Connect with us at firstname.lastname@example.org to learn more about how we can help.
The ProTrap XG device is optimized to process 50 µg of protein.
For reproducible and maximal protein precipitation, the maximum SDS content in your sample during precipitation should be 1%. If your extraction or lysis buffer contains more than 1% SDS, dilute it with a buffer containing a maximum ionic strength of 300 mM.
Spin speeds are based on a standard benchtop microcentrifuge with 24 x 1.5/2.0 mL rotor.
Times provided are guidelines only.
If more than a few microliters of liquid remains in the Filtration Cartridge after any spin, return it to the centrifuge and repeat the spin, or consider increasing the spin speed. 3000 ×g (6000 rpm) is recommended for subsequent spins and the ProTrap XG has been tested up to 9000 ×g (10,000 rpm).
All chemicals and reagents should be ACS grade/HPLC grade or better.
5 M NaCl in water
80% Formic Acid in water (chilled to –20°C)
TOP DOWN SAMPLE PREP PROTOCOL
For reproducible and maximal protein precipitation, the maximum SDS content in your sample during precipitation should be 1%. If your extraction or lysis buffer contains more than 1% SDS, dilute it with a buffer containing a maximum ionic strength of 300 mM. For the best experience, your sample should contain at least 50 mM NaCl. If you need to add NaCl, use a 5 M NaCl solution.
If further protein clean-up is desired, resolubilized proteins may also be subject to SPE using the provided OPTIONAL SPE Cartridge and the SPE Protein/Peptide Clean-up Protocol.