Anti-Histone H4 (acetyl K16) HIST1H4A Monoclonal Antibody

• Catalog Number: orb548501
• Category: Antibodies
• Description: Anti-Histone H4 (acetyl K16) HIST1H4A Monoclonal Antibody. Tested in WB, IHC, ICC/IF, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: COC-8
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human Histone H4 (acetyl K16)
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 11 kDa
• UniProt ID: P62805
• 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 H4; HIST1H4A; H4/A, H4FA; HIST1H4B; H4/I,
• Note: For research use only
• Application notes: WB 1:500-1:2000IHC 1:50-1:200ICC/IF 1:50-1:200FC 1:50. Restore with deionized water (or equivalent) for reconstitution volume of 1.0 mL
• Expiration Date: 12 months from date of receipt.

Immunofluorescent analysis of HeLa cells treated with TSA, using Histone H4 (acetyl K16) Antibody.

IHC analysis of HIST1H4A using anti-HIST1H4A antibody (orb548501). HIST1H4A was detected in paraffin-embedded 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-HIST1H4A Antibody (orb548501) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

Western blot analysis of HIST1H4A expression in HeLa cell lysate treated with TSA using anti-HIST1H4A antibody (orb548501). 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-HIST1H4A antigen affinity purified polyclonal antibody (Catalog # orb548501) at 0.5 ug/mL overnight at 4°C, 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.