Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 ug of lysates per lane. Antibodies: IL-21 41008 (1 ug/mL), IL-21 2463 (5 ug/mL), beta-actin (5 ug/mL) and beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human HL-60 Cell Lysate
Loading: 15 ug of lysates per lane. Antibodies: IL-21 41008 (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.(A) Absence of blocking peptide(B) Presence of blocking peptide.

Immunocytochemistry Validation of IL-21 in K562 Cells
Immunocytochemical analysis of K562 cells using anti-IL-21 antibody (41008) at 2 ug/mL. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Induced Expression Validation of IL-21 expresssion in patients with Crohn’s Disease (Monteleone et al, 2416)
Enhanced IL-21 was observed in involved but not uninvolvedCD, and was not associated with any CD phenotype, such as fibrostenosing disease. IL-21 expression was detected by anti-IL-21 antibodies (41008).