Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling Bcl G with ab184925 at 1/250 dilution, followed by Goat anti-rabbit I Alexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary) at 1/500 dilution (red).The negative controls are as follows:-ve control 1: ab184925 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.
All lanes : Anti-Bcl G antibody [EPR17666] (ab184925) at 1/2000 dilutionLane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysatesLane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysatesLane 3 : A549 (Human lung carcinoma) whole cell lysatesLane 4 : Human fetal liver lysatesLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Anti-Bcl G antibody [EPR17666] (ab184925) at 1/5000 dilution + Human testis lysates at 10 µgSecondaryGoat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
All lanes : Anti-Bcl G antibody [EPR17666] (ab184925) at 1/2000 dilutionLane 1 : Mouse brain lysatesLane 2 : Mouse heart lysatesLane 3 : Mouse kidney lysatesLane 4 : Mouse spleen lysatesLane 5 : Rat brain lysatesLane 6 : Rat heart lysatesLane 7 : C6 (Rat glial tumor cells) whole cell lysatesLane 8 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysatesLane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysatesLane 10 : 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 tonsil tissue labeling Bcl G with ab184925 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasm staining on lymphocytes of Human tonsil is observed. Counter stained with Hematoxylin.Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. J Biol Chem. 2001 Jan 26;276(4):2780-5.
Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix tissue labeling Bcl G with ab184925 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic staining on tumor cells of Human cervical squamous cell carcinoma is observed. Counter stained with Hematoxylin.Negative control: Using 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 Mouse kidney tissue labeling Bcl G with ab184925 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic staining on mouse kidney tubules is observed. Counter stained with Hematoxylin.Negative control: Using 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 Rat colon tissue labeling Bcl G with ab184925 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic staining on glandular epithelium of rat colon is observed. Counter stained with Hematoxylin.Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl G with ab184925 at 1/130 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.
Bcl G was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab184925 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184925 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: Rabbit monoclonal IgG (ab172730) instead of ab184925 in HeLa whole cell extract.Blocking and dilution buffer and concentration: 5% NFDM/TBST.