The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 5-10 µg/mg of lysate.
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notesIs unsuitable for WB.
FunctionPromotes cell death. Successfully competes for the binding to Bcl-X(L), Bcl-2 and Bcl-W, thereby affecting the level of heterodimerization of these proteins with BAX. Can reverse the death repressor activity of Bcl-X(L), but not that of Bcl-2 (By similarity). Appears to act as a link between growth factor receptor signaling and the apoptotic pathways.
Tissue specificityExpressed in a wide variety of tissues.
Sequence similaritiesBelongs to the Bcl-2 family.
DomainIntact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.
Post-translational modificationsPhosphorylated on one or more of Ser-75, Ser-99, Ser-118 and Ser-134 in response to survival stimuli, which blocks its pro-apoptotic activity. Phosphorylation on Ser-99 or Ser-75 promotes heterodimerization with 14-3-3 proteins. This interaction then facilitates the phosphorylation at Ser-118, a site within the BH3 motif, leading to the release of Bcl-X(L) and the promotion of cell survival. Ser-99 is the major site of AKT/PKB phosphorylation, Ser-118 the major site of protein kinase A (CAPK) phosphorylation. Ser-75 is phosphorylated by AKT/PKB, protein kinase A and PIM2.
Cellular localizationMitochondrion outer membrane. Cytoplasm. Upon phosphorylation, locates to the cytoplasm.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma (left) and mouse renal cell carcinoma (right) tissue labelling Bad with ab114105 at 1/5000 (0.2µg/ml) and 1/1000 (1µg/ml). Detection: DAB.
Detection of Human Bad by Western blot of Immunoprecipitate.
Bad was immunoprecipitated from HeLa whole cell lysate, using ab114105 at 10 µg/mg lysate (1 mg for IP, 20% of IP loaded). ab114105 was used for subsequent blotting at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.
References for Anti-Bad antibody (ab114105)
has not yet been referenced specifically in any publications.
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