All lanes : Anti-A RAF antibody [EPR16208] (ab200653) at 1/10000 dilutionLane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysateLane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
All lanes : Anti-A RAF antibody [EPR16208] (ab200653) at 1/1000 dilutionLane 1 : HT-29 (Human colorectal adenocarcinoma cells) whole cell lysateLane 2 : A375 (Human malignant melanoma) whole cell lysateLane 3 : Human fetal heart lysateLane 4 : Human fetal kidney lysateLane 5 : Human bladder lysateLysates/proteins at 10 µg per lane.SecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Anti-A RAF antibody [EPR16208] (ab200653) at 1/1000 dilution + Human fetal brain lysate at 10 µgSecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling A RAF with ab200653 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.Cytoplasm staining on Human cervix carcinoma tissue is observed.Counter stained with Hematoxylin.Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling A RAF with ab200653 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.Cytoplasm staining on Human kidney tissue is observed.Counter stained with Hematoxylin.Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling A RAF with ab200653 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Cytoplasm staining on HeLa cell line is observed.The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:-ve control 1: ab200653 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK293 (Human embryonic kidney) cells labeling A RAF with ab200653 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Cytoplasm staining on HEK293 cell line is observed.The nuclear counter stain is DAPI (blue).Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:-ve control 1: ab200653 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling A RAF with ab200653 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.