Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling MEK3 + MEK6 with ab181555 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:-1. ab181555 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/50000 dilutionLane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysatesLane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysatesLane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysatesLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/10000 dilution + Human MEK6 recombinant protein fragment at 10 µgSecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/5000 dilutionLane 1 : Rat liver lysatesLane 2 : Mouse liver lysatesLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/2000 dilutionLane 1 : Mouse heart lysatesLane 2 : Mouse kidney lysatesLane 3 : Rat heart lysatesLane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysatesLane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysatesLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Human skeletal muscle is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary ab.
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Mouse skeletal muscle is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary ab.
Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Rat skeletal muscle is observed. Counter stained with Hematoxylin.Negative control: Used PBS instead of primary ab.
MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab181555 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab33866 (Rabbit monoclonal [EP557Y] to MEK6) at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.Blocking and dilution buffer and concentration: 5% NFDM/TBST.