Histone H3.3 Antibody (YA367)

Histone H3.3 Antibody (YA367) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Histone H3.3.

Histone H3, a fundamental component of the nucleosome, serves as a linchpin in the intricate process of wrapping and compacting DNA into chromatin, which in turn restricts DNA accessibility to cellular machineries requiring DNA as a template. This histone, alongside its counterparts, assumes a pivotal role in pivotal cellular functions, including transcription regulation, DNA repair, DNA replication, and the maintenance of chromosomal stability. The regulation of DNA accessibility involves a sophisticated interplay of post-translational modifications collectively referred to as the histone code, coupled with the dynamic remodeling of nucleosomes. The nucleosome itself comprises a histone octamer, composed of two molecules each of H2A, H2B, H3, and H4, assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. This octamer efficiently wraps approximately 147 base pairs of DNA, exemplifying its indispensable role in organizing chromatin structure and facilitating crucial genomic processes. Additionally, Histone H3 interacts with various cellular components such as TONSL, CHAF1A, CHAF1B, MCM2, and DNAJC9, further contributing to its multifaceted functions.

Free

P84243

Predicted band size: 15 kDa; Observed band size: 15 kDa

Protein A affinity purified.

Nucleus, Chromosome.

Non-conjugated

Unmodified

AB_3102848

Epigenetics and Nuclear Signaling

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:1,000 ;ICC/IF: 1:50-1:200 ;IHC-P: 1:50-1:500

• 
  Western blot analysis of extracts from Hela(lane 2(20μg) , C2C12(lane 3(20μg) and C6(lane 4(20ug) using Histone H3.3(HY-P80177 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling Histone H3.3 with Histone H3.3 Antibody (HY-P80177) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Histone H3.3 Antibody (HY-P80177) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of Hela cells labeling Histone H3.3 with Histone H3.3 Antibody (HY-P80177) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Histone H3.3 Antibody (HY-P80177)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded mouse colon tissue using Histone H3.3 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80177, 1:200) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded mouse colon tissue using Histone H3.3 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80177, 1:200) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P, ChIP

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human Histone H3.3.AA range:50-100.