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ab7977 has been referenced in 17 publications.
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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.
All lanes : Anti-Bax antibody (ab7977) at 1/1000 dilution
Lane 1 : CHO whole cell lysate. Non-transfected.
Lane 2 : CHO whole cell lysate. Cell transfected with Bax-EGFP in pcDNA3.1
Lysates/proteins at 25 µg per lane.
Secondary
Goat anti-rabbit IRDye 800 at 1/30000 dilution
Performed 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 minutes
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.
This image is courtesy of an anonymous abreview.
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