All lanes : Anti-STARD4 antibody [EPR17847-32] (ab202060) at 1/1000 dilutionLane 1 : Human fetal liver lysateLane 2 : Human fetal heart lysateLysates/proteins at 10 µg per lane.SecondaryAnti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

All lanes : Anti-STARD4 antibody [EPR17847-32] (ab202060) at 1/1000 dilutionLane 1 : Mouse kidney lysateLane 2 : Mouse spleen lysateLane 3 : Rat kidney lysateLane 4 : Rat spleen lysateLane 5 : Rat liver lysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

All lanes : Anti-STARD4 antibody [EPR17847-32] (ab202060) at 1/1000 dilutionLane 1 : C6 (Rat glial tumor cells) whole cell lysateLane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysateLane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysateLane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysateLane 5 : Mouse liver 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 STRAD4 with ab202060 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic 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: ab202060 at 1/1000 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 SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling STRAD4 with ab202060 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on SH-SY5Y 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: ab202060 at 1/1000 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.

STARD4 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab202060 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab202060 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: NIH/3T3 whole cell lysate 10 µg (Input). Lane 2: ab202060 IP in NIH/3T3 whole cell lysate Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202060 in NIH/3T3 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 10 seconds.