H2AX phospho S139 Antibody

SKU: orb345732

Description

H2AX pS139 antibody

Images & Validation

• Tested Applications: ELISA, WB
• Dilution Range: ELISA: 1:5,000, WB: 1:1,000
• Reactivity: Human
• Application Notes:
  Anti-H2AX pS139 antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 15 kDa in size corresponding to phosphorylated H2AX protein by western blotting in the appropriate stimulated tissue or cell lysate or extract.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Anti-H2AX pS139 purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a phosphorylated synthetic peptide corresponding to the C-terminal region containing serine 139 of human H2AX protein.
• Purity: H2AX pS139 is directed against the phosphorylated form of human H2AX protein at the pS139 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity with non-phosphorylated human H2AX is minimal by ELISA and western blot. A BLAST analysis was used to suggest 100% cross reactivity with H2AX from human based on the sequence homology with the immunogen. Reactivity against homologues from other sources is not known.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.0 mg/mL
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

rabbit anti-H2AX pS139 antibody, rabbit anti-Histone H2AX pS139 antibody, H2AFX antibody, H2a/x, H2A.x, H2AX, histone H2A, gamma-H2AX, H2A histone family member X antibody

Western Blot of Rabbit Anti-H2AX pS139 Antibody. Lane 1: Opal Prestained Molecular Weight Marker. Lane 2: HEK293T Whole cell Lysate [+]. Lane 3: MCF-7 Whole cell Lysate (p/n orb348664) [+]. Lane 4: U-87-MG Whole cell Lysate (p/n orb692719) [-]. Primary Antibody: Anti-H2AXpS139 at 5 µg/ml overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n orb347654) at 1:40000 for 30 min at RT. Block: Universal Fluorescent Buffer (p/n orb348637) for 1hr at RT. Predicted MW: ~15kDa. Observed MW: ~17kDa. Exposure: 10sec.

Western Blot of Rabbit anti-H2AXpS139 antibody. Lane 1: HeLa Lysate stimulated with adriamycin (24 hr). Lane 2: HeLa Lysate unstimulated. Load: 35 µg per lane. Primary antibody: H2AXpS139 antibody at 1:1000 for overnight at 4°C. Secondary antibody: HRP rabbit secondary antibody at 1:10000 for 45 min at RT. Block: 5% BLOTTO overnight at 4°C. Predicted/Observed size: 15.1 kDa, ~18 kDa for H2AXpS139. Other band(s): none.

Quick Database Links

UniProt

P16104

RefSeq (Protein):NP_002096.1