Anti-Histone H3 (mono+di+tri methyl K79) HIST1H3A Rabbit Monoclonal Antibody

• Catalog Number: orb548491
• Category: Antibodies
• Description: Anti-Histone H3 (mono+di+tri methyl K79) HIST1H3A Rabbit Monoclonal Antibody. Tested in WB, IHC applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: H-8
• Tested applications: IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human Histone H3 (mono+di+tri methyl K79)
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 17 kDa
• UniProt ID: P68431
• 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.
• Alternative names: Histone H3.1; Histone H3/a; Histone H3/b; Histone
• Note: For research use only
• Application notes: WB 1:500-1:1000IHC 1:1000-1:2000. Restore with deionized water (or equivalent) for reconstitution volume of 1.0 mL
• Expiration Date: 12 months from date of receipt.

Immunohistochemical analysis of paraffin-embedded rat colon, using Histone H3 (mono+di+tri methyl K79) Antibody (orb548491) HIST1H3A was detected in paraffin-embedded tissue section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-HIST1H3A Antibody (orb548491) overnight at 4 Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

Western blot analysis of Histone H3 (mono+di+tri methyl K79) expression in (1) NIH/3T3 cell lysate; (2) A549 cell lysate (orb548491). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HIST1H3A monoclonal antibody (Catalog # orb548491) overnight at 4 then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for HIST1H3A.