Anti human lamp2a ab

Triton X-100 cell lysates and trichloroacetic acid–concentrated supernatants from monocytes were resolved by 12% SDS/PAGE and electrotransferred. Filters were probed with 3ZD anti-IL-1β mAb (IgG1; obtained from the National Cancer Institute Biological Resources Branch), anti-GSDMDC1 (Novus Biologicals), anti-human LAMP2A Ab (Abcam), anti human α-tubulin mAb (Sigma-Aldrich) and anti-GAPDH (Novus Biologicals), followed by the relevant secondary Ab (Dako) and developed with ECL-plus (BioRad) (GE Healthcare), as described^{9 (link),34 (link)}. The total amounts of cell culture supernatants and 10 μg of proteins of cell lysates (corresponding to about 25% of the total cell lysates) were loaded for IL-1β experiments. For GSDMD and LAMP2A detections 40 μg of cell lysate were loaded.

Monocytes were isolated from PBMCs using CD14 MicroBeads (Miltenyi Biotec) and activated with LPS or LRZ. Cytofluorimetry was performed as indicated^{57 (link)}. Cells were stained with anti-human LAMP2A Ab (Abcam) followed by the anti-rabbit FITC secondary Ab (Bethyl Laboratories, Inc). Samples were acquired using a CyAn flow cytometer (Beckman Coulter) and analyzed by the Summit V4.3 software (DAKO)^{32 (link)}.