Bad Rabbit Polyclonal Antibody

• Catalog Number: orb10165
• Category: Antibodies
• Description: Bad Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Bad (120-204/204aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=3ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 18 kDa
• Target: BAD
• UniProt ID: Q92934
• RRID: AB_10752941
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: BBC 2; BBC2; BBC6; Bcl 2 Antagonist of Cell Death;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (Black line): Jurkat (Black). Primary Antibody (green line): Rabbit Anti-Bad antibody (orb10165), Dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, Dilution: 1 µg/test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 15 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: A431 (Black). Primary Antibody (green line): Rabbit Anti-Bad antibody (orb10165), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at -20℃. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Bad) Polyclonal Antibody, Unconjugated (orb10165) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat stomach), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Bad) Polyclonal Antibody, Unconjugated (orb10165) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Protein: Brain (Mouse) Lysates, 30 ug, Primary: Anti-Bad (orb10165) at 1:200, Secondary: HRP conjugated Goat Anti-Rabbit IgG (orb572747) at 1: 5000, Predicted band size: 18 kD, Observed band size: 18 kD.

Sample: Hela Cell lysate 30 ug, Primary: Anti-Bad (orb10165) at 1:300, Secondary: HRP conjugated Goat-Anti-Rabbit IgG (orb572747) at 1:5000, Predicted band size:18 kD, Observed band size: 27 kD.

Sample: Raji Cell lysate 30 ug, Primary: Anti-Bad (orb10165) at 1:300, Secondary: HRP conjugated Goat-Anti-Rabbit IgG (orb572747) at 1:5000, Predicted band size:18 kD, Observed band size: 27 kD.

Tissue/cell: Human kidney tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37℃ for 20 min, Incubation: Anti-Bad Polyclonal Antibody, Unconjugated (orb10165) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.