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, untreated or treated with lipopolysaccharide (1 µg/ml) for 6 hours, labeling COX2 / Cyclooxygenase 2 with ab188184 at 1/150 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased cytoplasmic staining on RAW 264.7 cells treated with lipopolysaccharides (1 µg/ml) for 6 hours.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line, treated with 1μg/ml LPS for 6h (red) and untreated control (green), labeling COX2 / Cyclooxygenase 2 with ab188184 at 1/600 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
COX2 / Cyclooxygenase 2 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab188184 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab188184 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10μg (Input).
Lane 2: ab188184 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188184 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure: 30 seconds.
has not yet been referenced specifically in any publications.
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