Rabbit anti-Human Bad Polyclonal Antibody

Bad, phosphorylated (Ser99) (Bcl2 Antagonist Of Cell Death, BAD, Bcl-2-binding Component 6, Bcl-XL/Bcl-2-Associated Death Promoter, Bcl-2-like 8 Protein, BAD, BBC6, BCL2L8)

Cat. Num. AAA628799

• Reactivity: Human
• Applications: ELISA, Western Blot, Immunohistochemistry, Gel Super Shift Assay
• Purity: Affinity Purified; Purified by immunoaffinity chromatography.
• Synonyms: Bad; Polyclonal Antibody; phosphorylated (Ser99) (Bcl2 Antagonist Of Cell Death; BAD; Bcl-2-binding Component 6; Bcl-XL/Bcl-2-Associated Death Promoter; Bcl-2-like 8 Protein; BBC6; BCL2L8); Anti -Bad; anti-Bad antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: Recognizes human Bad when phosphoryated at Ser99..
• Purity/Purification: Affinity Purified; Purified by immunoaffinity chromatography.
• Form/Format: Supplied as a liquid in PBS, 0.09% sodium azide.

• Applicable Applications for anti-Bad antibody: ELISA (EL/EIA), Western Blot (WB), Immunohistochemistry (IHC), Gel Shift Assay (GS/EMSA)
• Application Notes: Suitable for use in ELISA, Western Blot, Dot Blot, and Immunohistochemistry.; Dilution: ELISA: 1:1,000; Western Blot: 1:100-1:500; Immunohistochemistry: 1:50-1:100; Dot Blot: 1:500
• Immunogen: Synthetic phosphopeptide corresponding to amino acid residues surrounding Ser99 of human Bad (KLH).
• Preparation and Storage: May be stored at 4 degree C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degree C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Immunofluorescence (IF)

(Fluorescent confocal image of HeLa cells stained with phospho-Bad-S99 antibody. HeLa cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with AP3039a phospho-Bad-S99 primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). Note the highly specific localization of the phospho-Bad-S99 mainly to the cytoplasm.)

Dot Blot (DB)

(Dot blot analysis of anti-hBad-pS99 Phospho-specific Pab on nitrocellulose membrane. 50ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5ug per ml.)

Immunohistochemistry (IHC)

(Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.)

Related Product Information for anti-Bad antibody

Bad is a member of the BCL-2 family. BCL-2 family members are known to be regulators of programmed cell death. This protein positively regulates cell apoptosis by forming heterodimers with BCL-xL and BCL-2, and reversing their death repressor activity. Proapoptotic activity of this protein is regulated through its phosphorylation. Protein kinases AKT and MAP kinase, as well as protein phosphatase calcineurin are found to be involved in the regulation of this protein. Bad is phosphorylated 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.

Product Categories/Family for anti-Bad antibody

Antibodies; Abs to Apoptosis