PRODUCT CODE: ET1608-9

Acetyl-Histone H3 (K56) Recombinant Rabbit Monoclonal Antibody [SU30-10] (ET1608-9)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Histone H3(acetyl K56) on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-9, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Histone H3(acetyl K56) on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-9, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Histone H3(acetyl K56) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-9, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Histone H3(acetyl K56) in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-9, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Histone H3(acetyl K56) in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-9, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Histone H3(acetyl K56) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9, 1/50) for 30 minutes at room temperature. 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 kidney tissue using anti-Histone H3(acetyl K56) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-9, 1/50) for 30 minutes at room temperature. 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.
Western blot analysis of Histone H3(acetyl K56) on HepG2 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1608-9, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Acetyl-Histone H3 (K56) Recombinant Rabbit Monoclonal Antibody [SU30-10] (ET1608-9)

Immunogen

Synthetic peptide within human histone h3 aa 31-80 / 136.

Host

Rabbit

Modification

Acetyl

Modification Site

K56

Positive Control

Hela, HepG2, A549, human kidney tissue, mouse kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SU30-10

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

15 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Acetyl-Histone H3 (K56)

SYNONYMS

H3.3A antibody; HIST1 cluster, H3E antibody; H3 histone family, member A antibody; H3.1 antibody; H3/l antibody; H3F3 antibody; H3FF antibody; H3FJ antibody; H3FL antibody; Histone gene cluster 1, H3 histone family, member E antibody; histone H3.1t antibody; Histone H3/o antibody; FLJ92264 antibody; H 3 antibody; H3 antibody; H3 histone family, member B antibody; H3 histone family, member C antibody; H3 histone family, member D antibody; H3 histone family, member F antibody; H3 histone family, member H antibody; H3 histone family, member I antibody; H3 histone family, member J antibody; H3 histone family, member K antibody; H3 histone family, member L antibody; H3 histone family, member T antibody; H3 histone, family 3A antibody; H3/A antibody; H3/b antibody; H3/c antibody; H3/d antibody; h3/f antibody; H3/h antibody; H3/i antibody; H3/j antibody; H3/k antibody; H3/t antibody; H31_HUMAN antibody; H3F1K antibody; H3F3A antibody; H3FA antibody; H3FB antibody; H3FC antibody; H3FD antibody; H3FH antibody; H3FI antibody; H3FK antibody; HIST1 cluster, H3A antibody; HIST1 cluster, H3B antibody; HIST1 cluster, H3C antibody; HIST1 cluster, H3D antibody; HIST1 cluster, H3F antibody; HIST1 cluster, H3G antibody; HIST1 cluster, H3H antibody; HIST1 cluster, H3I antibody; HIST1 cluster, H3J antibody; HIST1H3A antibody; HIST1H3B antibody; HIST1H3C antibody; HIST1H3D antibody; HIST1H3E antibody; HIST1H3F antibody; HIST1H3G antibody; HIST1H3H antibody; HIST1H3I antibody; HIST1H3J antibody; HIST3H3 antibody; Histone 1, H3a antibody; Histone 1, H3b antibody; Histone 1, H3c antibody; Histone 1, H3d antibody; Histone 1, H3e antibody; Histone 1, H3f antibody; Histone 1, H3g antibody; Histone 1, H3h antibody; Histone 1, H3i antibody; Histone 3, H3 antibody; histone cluster 1 H3 family member a antibody; histone cluster 1 H3 family member b antibody; histone cluster 1 H3 family member c antibody; histone cluster 1 H3 family member d antibody; histone cluster 1 H3 family member e antibody; histone cluster 1 H3 family member f antibody; histone cluster 1 H3 family member g antibody; histone cluster 1 H3 family member h antibody; histone cluster 1 H3 family member i antibody; histone cluster 1 H3 family member j antibody; Histone cluster 1, H3a antibody; Histone cluster 1, H3b antibody; Histone cluster 1, H3c antibody; Histone cluster 1, H3d antibody; Histone cluster 1, H3e antibody; Histone cluster 1, H3f antibody; Histone cluster 1, H3g antibody; Histone cluster 1, H3i antibody; Histone cluster 1, H3j antibody; Histone gene cluster 1, H3 histone family, member A antibody; Histone gene cluster 1, H3 histone family, member B antibody; Histone gene cluster 1, H3 histone family, member C antibody; Histone gene cluster 1, H3 histone family, member D antibody; Histone gene cluster 1, H3 histone family, member F antibody; Histone gene cluster 1, H3 histone family, member G antibody; Histone gene cluster 1, H3 histone family, member H antibody; Histone gene cluster 1, H3 histone family, member I antibody; Histone gene cluster 1, H3 histone family, member J antibody; Histone gene cluster 1, H3A antibody; Histone gene cluster 1, H3B antibody; Histone gene cluster 1, H3C antibody; Histone gene cluster 1, H3D antibody; Histone gene cluster 1, H3E antibody; Histone gene cluster 1, H3F antibody; Histone gene cluster 1, H3G antibody; Histone gene cluster 1, H3H antibody; Histone gene cluster 1, H3I antibody; Histone gene cluster 1, H3J antibody; Histone H 3 antibody; Histone H3.1 antibody; Histone H3.2 antibody; Histone H3.3 antibody; Histone H3/a antibody; Histone H3/b antibody; Histone H3/c antibody; Histone H3/d antibody; Histone H3/f antibody; Histone H3/h antibody; Histone H3/i antibody; Histone H3/j antibody; Histone H3/k antibody; Histone H3/l antibody; Histone H3/m antibody

SEQUENCE SIMILARITIES

Belongs to the histone H3 family.

DEVELOPMENTAL STAGE

Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

POST-TRANSLATIONAL MODIFICATION

Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me). Acetylation at Lys-123 (H3K122ac) by EP300/p300 plays a central role in chromatin structure: localizes at the surface of the histone octamer and stimulates transcription, possibly by promoting nucleosome instability.; Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.; Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.; Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin. Monomethylation at Lys-57 (H3K56me1) by EHMT2/G9A in G1 phase promotes interaction with PCNA and is required for DNA replication.; Phosphorylated at Thr-4 (H3T3ph) by HASPIN during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MAP3K20 isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Thr-12 (H3T11ph) by chromatin-associated CHEK1 regulates the transcription of cell cycle regulatory genes by modulating acetylation of Lys-10 (H3K9ac). Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.; Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.; Lysine deamination at Lys-5 (H3K4all) to form allysine is mediated by LOXL2. Allysine formation by LOXL2 only takes place on H3K4me3 and results in gene repression.; Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.; Butyrylation of histones marks active promoters and competes with histone acetylation. It is present during late spermatogenesis.; Succinylation at Lys-80 (H3K79succ) by KAT2A takes place with a maximum frequency around the transcription start sites of genes. It gives a specific tag for epigenetic transcription activation. Desuccinylation at Lys-123 (H3K122succ) by SIRT7 in response to DNA damage promotes chromatin condensation and double-strand breaks (DSBs) repair.; Serine ADP-ribosylation constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage. Serine ADP-ribosylation at Ser-11 (H3S10ADPr) is mutually exclusive with phosphorylation at Ser-11 (H3S10ph) and impairs acetylation at Lys-10 (H3K9ac).

SUBCELLULAR LOCATION

Nucleus, Chromosome.

FUNCTION

Eukaryotic histones are basic and water soluble nuclear proteins that form hetero-octameric nucleosome particles by wrapping 146 base pairs of DNA in a left-handed super-helical turn sequentially to form chromosomal fiber. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form the octamer; formed of two H2A-H2B dimers and two H3-H4 dimers, forming two nearly symmetrical halves by tertiary structure. Over 80% of nucleosomes contain the linker Histone H1, derived from an intronless gene, that interacts with linker DNA between nucleosomes and mediates compaction into higher order chromatin. Histones are subject to posttranslational modification by enzymes primarily on their N-terminal tails, but also in their globular domains. Such modifications include methylation, citrullination, acetylation, phosphorylation, sumoylation, ubiquitination and ADP-ribosylation.

CITATIONS

  • Qin, Ge et al.

    Panobinostat (LBH589) inhibits Wnt/β-catenin signaling pathway via upregulating APCL expression in breast cancer. | Cellular Signalling [2019]