Histone H4 (acetyl K5) HIST1H4A Monoclonal Antibody

• Catalog Number: orb548502
• Category: Antibodies
• Description: Histone H4 (acetyl K5) HIST1H4A Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: CCA-8
• Tested applications: ICC, IF, IHC, IP, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human Histone H4 (acetyl K5) Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Dilution range: WB 1:1000-1:2000IHC 1:50-1:200ICC/IF 1:50-1:200IP 1:50
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• UniProt ID: P62805
• Storage: Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

IHC analysis of HIST1H4A using anti-HIST1H4A antibody (orb548502). 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 (orb548502) overnight at 4°C. Biotinylated goat anti Rabbit IgG IgG antibody 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 TSA treated HeLa cell lysate using anti-HIST1H4A antibody (orb548502). 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 # orb548502) 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 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 # orb90444) with Tanon 5200 system. A specific band was detected for HIST1H4A.