Hrp conjugated anti mouse and anti rabbit igg antibodies

Anti-Cullin1 and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were purchased from Calbiochem (La Jolla, CA). Anti-EF1α, anti-Skp1, and anti-ubiquitin antibodies were obtained from Millipore (Billerica, MA). The Anti-flag tag monoclonal antibody was bought from Sigma chemicals (St. Louis, MO). HRP-conjugated anti-mouse and anti-rabbit IgG antibodies for immunoblot assays were obtained from Santa Cruz Biotech Inc. (Santa Cruz, CA). AlexaFluor488- and AlexaFluor594-conjuagted anti-mouse and anti-rabbit IgG antibodies were purchased from Molecular Probes (Invitrogen) for immunofluorescence assays. Human pooled sera were prepared from 10 scrub typhus patients (IFA titer≥1∶1280) and 10 healthy volunteers following institutional review board approval and the receipt of informed consent from all subjects.

BJ-5ta cells were cultured in a DMEM and Medium199 combination, whereas HaCaT cells were grown in DMEM. Recombinant human TNF-α, IFN-γ, and ELISA kits for IL-8, IL-6, MDC, and RANTES are from Biolegend (San Diego, CA, USA). Sigma-Aldrich was used to obtain all chemical reagents (St. Louis, MO, USA). Santa Cruz Biotechnology supplied the ICAM-1, Actin, PCNA, HRP-conjugated anti-mouse, and anti-rabbit IgG antibodies (Dallas, TX, USA). Cell Signaling Technologies sold the researchers antibodies against p-IκBα, IκBα, p65, p-JAK2, p-STAT1, p-STAT3, p-p38, p38, p-JNK, and JNK (Danvers, MA, USA).