PRODUCT CODE: R1110-1

Mono-Methyl-Histone H3 (Lys4) Rabbit Polyclonal Antibody (R1110-1)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of Mono-mehtyl-Histone H3(Lys4) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (R1110-1, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control:    <br />
A: NCCIT    <br />
B: purified 293T Histone   <br />
C: F9        <br />
D: F9 with Histone H3 peptide-unmodifed  <br />
E: F9 with Histone H3 peptide-mono methyl K4   lysates using anti-Histone H3(mono methyl K4) polyclonal antibody.
  • Western blot analysis of Mono-mehtyl-Histone H3(Lys4) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (R1110-1, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control:    <br />
A: NCCIT    <br />
B: purified 293T Histone   <br />
C: F9        <br />
D: F9 with Histone H3 peptide-unmodifed  <br />
E: F9 with Histone H3 peptide-mono methyl K4   lysates using anti-Histone H3(mono methyl K4) polyclonal antibody.
  • ICC staining of Mono-mehtyl-Histone H3(Lys4) in NCCIT 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 (R1110-1, 1/100) 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 liver carcinoma tissue using anti-Mono-mehtyl-Histone H3(Lys4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-1, 1/200)  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 human colon carcinoma tissue using anti-Mono-mehtyl-Histone H3(Lys4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-1, 1/200)  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 human pancreas tissue using anti-Mono-mehtyl-Histone H3(Lys4) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-1, 1/200)  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.
  • Flow cytometric analysis of Mono-mehtyl-Histone H3(Lys4) was done on SHSY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (R1110-1, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Mono-mehtyl-Histone H3(Lys4) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (R1110-1, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
A: NCCIT
B: purified 293T Histone
C: F9
D: F9 with Histone H3 peptide-unmodifed
E: F9 with Histone H3 peptide-mono methyl K4 lysates using anti-Histone H3(mono methyl K4) polyclonal antibody.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

Mono-Methyl-Histone H3 (Lys4) Rabbit Polyclonal Antibody (R1110-1)

Immunogen

Peptide

Host

Rabbit

Modification

Mono-Methyl

Modification Site

Lys4

Positive Control

NCCIT, human liver carcinoma tissue, human colon carcinoma tissue, human pancreas tissue, SHSY5Y.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Immunogen affinity purified

MOLECULAR WEIGHT

17 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1

  • ICC

  • 1:50

TARGET

UNIPROT #

PROTEIN NAME

Mono-Methyl-Histone H3 (Lys4)

SYNONYMS

HIST1 cluster, H3J antibody; Histone gene cluster 1, H3 histone family, member E antibody; Histone gene cluster 1, H3G antibody; H3 histone family, member J antibody; HIST1 cluster, H3E antibody; HIST1 cluster, H3I antibody; Histone gene cluster 1, H3C antibody; FLJ92264 antibody; H3 histone family, member A 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 K antibody; H3 histone family, member L antibody; H3 histone family, member T antibody; H3 histone, family 3A antibody; H3.1 antibody; H3.3A 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/l antibody; H3/t antibody; H31_HUMAN antibody; H3F1K antibody; H3F3 antibody; H3F3A antibody; H3FA antibody; H3FB antibody; H3FC antibody; H3FD antibody; H3FF antibody; H3FH antibody; H3FI antibody; H3FJ antibody; H3FK antibody; H3FL 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; 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 1, H3j 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, H3D antibody; Histone gene cluster 1, H3E antibody; Histone gene cluster 1, H3F 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 H3.1t antibody; Histone H3.2 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; Histone H3/o 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

FUNCTION

The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. 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.