Dinitrophenyl coupled human serum albumin dnp hsa

Cell culture medium (RPMI 1640 medium) was purchased from Invitrogen (Carlsbad, CA), fetal bovine serum (FBS) from Quality Biological, Inc. (Gaithersburg, MD). Recombinant mouse IL-3 and SCF were purchased from Shenandoah Biotechnology, Inc (Warwick, PA). Purified IgE (clone c38–2, κ isotype) was purchased from BD Biosciences (San Diego, CA). Dinitrophenyl-coupled human serum albumin (DNP-HSA) and histamine were purchased from Sigma-Aldrich (St. Louis, MO). Dr. Daniel Conrad (VCU) generously provided mouse anti-DNP IgE for in vivo experiments. Antibodies against c-Kit (PE anti-mouse CD117) and FcεRI (APC anti-mouse FcεRIα) were purchased from Biolegend (San Diego, CA) and used at a dilution of 1:100.

Recombinant mouse IL-3, SCF and IL-10 were purchased from Biolegend (San Diego, CA). Purified mouse IgE (clone C38-2, κ isotype) was purchased from BD Biosciences (Pharmingen division, San Diego, CA). Dinitrophenyl-coupled human serum albumin (DNP-HSA) and LPS (Cat# L-6529) were purchased from Sigma-Aldrich (St. Louis, MO). Histamine was purchased from Enzo Life Sciences (Farmingdale, NY). All western blotting antibodies were purchased from Cell Signaling Technology (Danvers, MA). Dr. Daniel Conrad (VCU) generously provided purified mouse anti-DNP IgE for in vivo experiments. Antibodies for c-Kit (PE anti-mouse CD117) and FcεRI (APC anti-mouse FcεRIα) were purchased from Biolegend (San Diego, CA) and used at a concentration 1:100. LPS levels were tested using Toxin Sensor^™ Chromogenic LAL Endotoxin Assay Kit from GenScript (Piscataway, NJ). IL-10 resuspended in PBS and PBS alone used in in vivo studies had LPS content of <0.1 EU/mol. Media and IL-10 resuspended in media for in vitro studies had LPS content of ~1.0 EU/mol. There was no significant difference between IL-10 and the respective vehicle control used when comparing LPS levels using unpaired Student’s t-test.