IL-6 was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate with ab179570 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab179570 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: RAW 264.7 treated with 100ng/ml lipopolysaccharides (LPS) for 3h, then together with 300ng/ml BFA for another 3h whole cell lysate, 10 µg (Input).
Lane 2: ab179570 IP in RAW 264.7 treated with 100ng/ml lipopolysaccharides (LPS) for 3h, then together with 300ng/ml BFA for another 3h whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab179570 in RAW 264.7 treated with 100ng/ml lipopolysaccharides (LPS) for 3h, then together with 300ng/ml BFA for another 3h whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling IL-6 with ab179570 at 1/100 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing an elevated cytoplasmic staining in RAW 264.7 cells treated with LPS (100 ng/ml, 6 h) then together with 300 ng/ml BFA for another 3h. The nuclear counter stain is DAPI (blue). Tubuin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.