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Anti-BAD Antibody Y208
|A synthetic peptide corresponding to residues in the N-term of human Bad was used as immunogen. The antibody does not cross-react with other Bcl-2 members.|
|Human, Mouse, Rat
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:2000 - 1:5000; IHC-P: Use at an assay dependent dilution; ICC/IF: 1:250 - 1:500; FC: 1:20 - 1:50; IP: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Protein A purified (Protein A or G Sepharose)
|Homo sapiens BCL2-associated agonist of cell death (BAD), transcript variant 2|
|Bad is a pro-apoptotic member of the Bcl-2 family. Bad will bind preferentially Bcl-xL over Bcl-2 through its partial (9-16 a.a) BH3 domain. Additionally, Bad can reverse the death repressor activity of Bcl-xL but not of Bcl-2. Upon dephosphorylation, activated Bad translocates to the mitochondria and displace Bcl-2 from tBid sequestration to initiate mitochondrial dysfunction. The switching on/off of its phosphorylation by growth/survival factors regulates Bad activity. |
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Western blot - Bad antibody [Y208]; Anti-Bad [Y208] antibody at 1/5000 dilution + Hela cell lysate.Predicted band size : 18 kDa.Observed band size : 23 kDa .
Immunohistochemistry (Paraffin-embedded sections) - Bad antibody [Y208]; Immunohistochemical staining of paraffin-embedded human lymphoma using ab32445 at 1/100 dilution.
Immunocytochemistry/ Immunofluorescence-Bad antibody [Y208](ab32445); ICC/IF image of ab32445 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor? 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry-Bad antibody [Y208](ab32445); Overlay histogram showing MCF-7 cells stained with ab32445 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.