1X10^6 unstimulated or PMA and ionomycin stimulated (in the presence of protein transport inhibitors) mouse Th2-polarized splenocytes were surface stained with 0.2 ug PE Anti-Mouse CD4 (GK1.5) (PE-65104, Clone: GK1.5). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C), and then intracellularly stained with 0.5 ug APC Anti-Mouse IL-4 (APC-65074, Clone:11B11).

1X10^6 unstimulated or PMA and ionomycin stimulated (in the presence of protein transport inhibitors) mouse Th2-polarized splenocytes were surface stained with 0.25 ug CoraLite® Plus 647 Anti-Mouse CD4 (GK1.5) (CL647-65104, Clone: GK1.5). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C), and then intracellularly stained with 0.25 ug CoraLite® Plus 488 Anti-Mouse IL-4 (CL488-65074, Clone:11B11).

1X10^6 Ramos cells were intracellularly stained with 0.8 ug CoraLite® Plus 488 Anti-Human IL-4 (CL488-66142, Clone:1G2C5) (red), or 0.8 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

1X10^6 Ramos cells were intracellularly stained with 0.4 ug CoraLite®594 Anti-Human IL-4 (CL594-66142, Clone:1G2C5) (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Purity of recombinant human IL-4 was determined by SDS- polyacrylamide gel electrophoresis. The protein was resolved in an SDS- polyacrylamide gel in reducing and non-reducing conditions and stained using Coomassie blue.

Recombinant human IL-4 (HZ-1004) stimulates dose-dependent proliferation of the TF-1 human erythroleukemic indicator cell line. Cell number was quantitatively assessed by PrestoBlue® cell viability reagent. TF-1 cells were treated with increasing concentrations of recombinant IL-4 for 72 hours. The EC50 was determined using a 4-parameter non-linear regression model. The EC50 range is 0.07-0.4 ng/mL.

Jurkat cells were subjected to SDS PAGE followed by western blot with 66142-1-Ig (IL-4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using IL-4 antibody (66142-1-Ig, Clone: 1G2C5 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66142-1-Ig (IL-4 antibody at dilution of 1:50 (under 10x lens).

Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 66142-1-Ig (IL-4 antibody at dilution of 1:50 (under 40x lens).

1X10^6 Ramos cells were intracellularly stained with 0.4 ug Anti-Human IL-4 (66142-1-Ig, Clone:1G2C5) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2a Isotype Control (66360-2-Ig, Clone: K11A1B2A2) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Standard curve of AuthentiKine™ Human IL-4 ELISA Kit KE00232.