Capillary-Based Immunoassay for Protein Expression

A capillary-based immunoassay was performed using the Size Separation Master Kit (SM-W004, ProteinSimple, San Francisco, CA, USA) with Split Buffer (12–230 kDa) by the Wes (WS3312, ProteinSimple, San Francisco, CA, USA) [47 (link),48 (link),49 (link),50 (link)]. Protein expression was measured by chemiluminescence and quantified as area under the curve using the Compass for Simple Western program (Compass for SW 5.0.1, ProteinSimple, San Francisco, CA, USA). Proteins were detected with the following primary antibodies: NF-kB p65 (8242, cell signaling); HMGB1 (D3E5) (6893, Cell signaling, Boston, MA, USA); NLRP3 (D4D8T) (15101S, cell signaling, Boston, MA, USA); ASC/TMS1 (D2W8U) (67924, Cell signaling, Boston, MA, USA); GAPDH (5174, Cell signaling, Boston, MA, USA).