Human Anti-IL21R Recombinant Antibody (clone ATR-107) (CAT#: FAMAB-1203CQ)

Figure 1 Total and phospho-STAT3 was detected by Western blot.

Purified T cells from healthy donorperipheral blood were pre-incubated with ATR-107 at concentrations from 0.03 ng/ml to 100 ng/ml or control antibody for 30 minutes, followed by stimulation with IL-21 (10 ng/ml) for an additional 15 minutes.

Zhu, M., Pleasic-Williams, S., Lin, T. H., Wunderlich, D. A., Cheng, J. B., & Masferrer, J. L. (2013). pSTAT3: a target biomarker to study the pharmacology of the anti-IL-21R antibody ATR-107 in human whole blood. Journal of translational medicine, 11(1), 65.

Figure 2 Healthy donorwhole blood was pretreated with ATR-107 or control Ab for 30 minutes prior to stimulation with 10 ng/ml IL-21 for 15 minutes.

Cells were fixed, permeabilized, stained with A647-labeled anti-phospho-STAT-3 antibody, and analyzed using flow cytometry gated on CD4+T cells. Representative histogram showing the baseline (gray area), IL-21 + control Ab (red line) and IL-21 with various amount of ATR-107.

Zhu, M., Pleasic-Williams, S., Lin, T. H., Wunderlich, D. A., Cheng, J. B., & Masferrer, J. L. (2013). pSTAT3: a target biomarker to study the pharmacology of the anti-IL-21R antibody ATR-107 in human whole blood. Journal of translational medicine, 11(1), 65.

Figure 3 ATR-107 concentration-dependently inhibited IL-21 induced STAT3 phosphorylation.

Inhibition of ATR-107 on pSTAT3 mean fluorescence intensity (MFI) induced by 10 ng/ml IL-21(n = 6).

Zhu, M., Pleasic-Williams, S., Lin, T. H., Wunderlich, D. A., Cheng, J. B., & Masferrer, J. L. (2013). pSTAT3: a target biomarker to study the pharmacology of the anti-IL-21R antibody ATR-107 in human whole blood. Journal of translational medicine, 11(1), 65.

Figure 4 ATR-107 competes with IL-21 for receptor binding on naïve B-cells (CD19+CD27-).

Representative flow cytometry histograms showing the overlay of thereceptor occupancy performed without (blue line), or with either excessive concentration of IL-21 (707 nM, red line) or ATR-107 (363 ug/ml, blackline). At least 2000 CD19+CD27-events were acquired for analysis.

Zhu, M., Pleasic-Williams, S., Lin, T. H., Wunderlich, D. A., Cheng, J. B., & Masferrer, J. L. (2013). pSTAT3: a target biomarker to study the pharmacology of the anti-IL-21R antibody ATR-107 in human whole blood. Journal of translational medicine, 11(1), 65.

Figure 1 Human Anti-IL21R Recombinant Antibody (FAMAB-1203CQ) in ELISA

ELISA analysis of FAMAB-1203CQ was performed by coating with human IL21R protein (His tag). Then blocked with BSA and incubated with FAMAB-1203CQ. The HRP-conjugated goat anti-IgG was used as a secondary antibody (1: 2000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.

Figure 2 Human Anti-IL21R Recombinant Antibody (FAMAB-1203CQ) in WB

Western blot analysis of FAMAB-1203CQ was performed by loading human IL21R protein onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with FAMAB-1203CQ and a HRP goat anti-IgG (H+L) was used as the secondary antibody (1: 3000). Chemiluminescent detection was performed.

Figure 3 Human Anti-IL21R Recombinant Antibody (FAMAB-1203CQ) in DB

Dot Blot analysis of FAMAB-1203CQ was performed by coating with human IL21R protein (His tag).
FAMAB-1203CQ incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-IgG (H+L) secondary antibody (1: 3000)
NC: Negative control, PC: Positive control