ab7977 staining Bax in SK-N-SH cells treated with isobavachalcone (ab141168), by ICC/IF. Increase of Bax expression correlates with increased concentration of isobavachalcone, as described in literature.The cells were incubated at 37°C for 6h in media containing different concentrations of ab141168 (isobavachalcone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab7977 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab7977 staining BAX in RAW 264.7 cells treated with etomidate (ab120311), by ICC/IF. Increase in BAX expression correlates with increased concentration of etomidate, as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab120311 (etomidate) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab7977 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
All lanes : Anti-Bax antibody (ab7977) at 1/1000 dilutionLane 1 : Mouse Heart tissue lysateLane 2 : Mouse Heart tissue lysateLysates/proteins at 20 µg per lane.SecondaryDonkey Anti-Rabbit IgG H&L (HRP) (ab16284) at 1/10000 dilution (HRP-conjugated Donkey anti-rabbit IgG polyclonal)developed using the ECL techniquePerformed under reducing conditions.Observed band size : 26 kDa (why is the actual band size different from the predicted?)Exposure time : 1 minuteThis image is courtesy of an anonymous AbreviewSee Abreview
ab7977 (1/1000) staining Bax in HeLa whole cel lysate ab7898.
All lanes : Anti-Bax antibody (ab7977) at 1/1000 dilutionLane 1 : CHO whole cell lysate. Non-transfected.Lane 2 : CHO whole cell lysate. Cell transfected with Bax-EGFP in pcDNA3.1Lysates/proteins at 25 µg per lane.SecondaryGoat anti-rabbit IRDye 800 at 1/30000 dilutionPerformed under reducing conditions.Observed band size : 48 kDa (why is the actual band size different from the predicted?)Additional bands at : 20 kDa,25 kDa. We are unsure as to the identity of these extra bands.Exposure time : 3 minutesThis image is courtesy of an anonymous abreview.Bax fused to EGFP should correspond to approximately 48 kDa as EGFP is 28 kDa and BAX is 21 kDa. 8% gel run under denaturing conditions.Blocked using 3% BSA for 2 hours at 25°C.Detection method: Infrared.Cells were transfected using Lipofectamine 2000 and harvested after ~18 hours.See Abreview
Immunohistochemical analysis of murine testis tissue sections, staining Bax with ab7977.Tissue was fixed with formaldehyde and blocked with 3% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 12 hours at 4°C. An undiluted HRP-conjugated anti-rabbit polyclonal IgG was used as the secondary antibody.See Abreview
ab7977 staining Human normal bladder. Staining is localized to cytoplasmic compartment.Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.