Phospho-H2AX (Ser139) Rabbit Polyclonal Antibody

• Catalog Number: orb6153
• Category: Antibodies
• Description: Phospho-H2AX (Ser139) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human Histone H2AX around the phosphorylation site of Tyr139 QA(p-S)QE
• 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=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 16 kDa
• Target: H2AX
• UniProt ID: P16104
• RRID: AB_10929309
• 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: phospho-Histone H2A.X(Ser140); H2AX_HUMAN; 2A.X va
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody (orb6153), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 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: Molt4. Primary Antibody (green line): Rabbit Anti-Phospho-Histone H2A.X (Ser139) antibody, dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-APC, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 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.

Sample: Lane 1: Mouse BV2 cell lysates, Lane 2: Rat Testis tissue lysates, Lane 3: Human 293T cell lysates, Lane 4: Human Jurkat cell lysates, Primary: Anti-Phospho-H2AX (Ser139) (orb6153) at 1/5000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 16 kDa, Observed band size: 15 kDa.

Tissue/Cell: human bladder carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Phospho-Histone H2A.X (Ser139) Polyclonal Antibody, Unconjugated (orb6153) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.