IL-10 Antibody (JE59-71)

Western Blot:IL-10 Antibody (JE59-71)-NBP3-32465-

Western Blot:IL-10 Antibody (JE59-71)-NBP3-32465-Analysis of IL-10 on different lysates with Rabbit anti-IL-10 antibody at 1/1,000 dilution.Lane 1: THP-1 cell lysate (30 ug/Lane)Lane 2: THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours cell lysate (30 µg/Lane)Lane 3: Mouse spleen tissue lysate (40 ug/Lane)Lane 4: Rat spleen tissue lysate (40 ug/Lane)Predicted band size: 21 kDaObserved band size: 21 kDaExposure time: 1 minutes 59 seconds;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

ICC/IF:IL-10 Antibody (JE59-71)-NBP3-32465-

ICC/IF:IL-10 Antibody (JE59-71)-NBP3-32465-Analysis of THP-1 cells treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours labeling IL-10 with Rabbit anti-IL-10 antibody at 1/500 dilution.Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IL-10 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594,) was used as the secondary antibody at 1/1,000 dilution.

Immunohistochemistry-Paraffin:IL-10 Antibody (JE59-71)-NBP3-32465-

Immunohistochemistry-Paraffin:IL-10 Antibody (JE59-71)-NBP3-32465-Rat spleen tissue with Rabbit anti-IL-10 antibody at 1/1,000 dilution.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.