All lanes : Anti-TIMM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilutionLane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) cell lysateLane 2 : HepG2 (Human liver hepatocellular carcinoma) cell lysateLane 3 : Daudi (Human Burkitt's lymphoma cell line) cell lysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Anti-TIMM44 antibody [EPR16821-26] (ab201453) at 1/10000 dilution + Human fetal liver lysate at 10 µgSecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
All lanes : Anti-TIMM44 antibody [EPR16821-26] (ab201453) at 1/2000 dilutionLane 1 : C6 (Rat glial tumor cells) cell lysateLane 2 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling TIMM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on HepG2 cell line. 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: ab201453 at 1/500 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 K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling TIMM44 with ab201453 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasm staining on K562 cell line. 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: ab201453 at 1/500 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.
TIMM44 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab201453 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab201453 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1500 dilution.Lane 1: K562 whole cell lysate 10 µg (Input). Lane 2: ab201453 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201453 in K562 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 1 second.