Immunofluorescence analysis of BID [cleavage site 59/60] was done on 70% confluent log phase A549. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with BID [cleavage site 59/60] Rabbit polyclonal Antibody (44436G) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized peptide derived from the 15 kDa caspase-8 cleavage product of mouse BID.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
Bid is a death agonist that heterodimerizes with either agonist BAX or antagonist BCL2. The encoded protein is a member of the BCL-2 family of cell death regulators. Bid induces ICE-like proteases and apoptosis. It is a mediator of mitochondrial damage induced by caspase-8 (CASP8); CASP8 cleaves this encoded protein, and the major proteolytic product p15 Bid translocates to mitochondria where it triggers cytochrome c release.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Cisplatin-induced apoptosis in auditory cells: role of death receptor and mitochondrial pathways.||Devarajan P,Savoca M,Castaneda MP,Park MS,Esteban-Cruciani N,Kalinec G,Kalinec F||Hearing research (174:45)||2002|