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Instrument was calibrated externally with a PepMix calibration kit (Bruker Daltonics).
The monitor was calibrated using an Eye-One calibration kit (X-Rite, Tewksbury, MA, USA).
For peptides and proteins, a ProteoMass™ Peptide and Protein MALDI-MS Calibration Kit was used to calibrate.
The instrument was calibrated externally using ProteoMass Peptide Calibration kit (Sigma), low mass gate value of 700 was selected.
The SEC column was calibrated using the Gel Filtration Calibration Kit (GE Healthcare) according to the manufacturer's protocol to detect the oligomerization state of the NLRP4 PYD.
A Superdex 200 gel filtration, calibrated using a Gel Filtration Calibration Kit LMW (GE Healthcare), was used to estimate the molecular weight of the native enzyme as 95 kDa, suggesting the enzyme exists as a homodimer.
Molecular weights were calibrated with polystyrene standards from Varian (Calibration Kit S-M2-10, AgilenTechnologieses, Palo Alto, CA, USA).
All fluorescence values were calibrated using the fura 2/AM Calcium Imaging Calibration kit (Invitrogen).
The software was calibrated using the InSpeck fluorescence Image Intensity Calibration Kit (6 :m beads; Molecular Probes, Eugene OR).
The gel filtration column was calibrated by using a low-molecular-weight calibration kit (GE Healthcare) according to the manufacturer's guidelines, allowing determination of the molecular mass of eluted proteins.
The column was calibrated using the gel filtration low-molecular-weight calibration kit (GE Healthcare).
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