Histone H3 (mono methyl K4) Rabbit Polyclonal Antibody
cat.: R1110-1
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, Dot Blot
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 15 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Histone H3 aa 1-49/136.
Positive control: NCCIT, PC-12, MCF-7, human liver carcinoma tissue, human colon carcinoma tissue, human pancreas tissue, SHSY5Y.
Subcellular location: Nucleus
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  Dot Blot
1:500-1:2000
1:50-1:200
1:50-1:200
1:50-1:100
1:1,000
Uniprot #: SwissProt: P68431 Human | P68433 Mouse | Q497B8 Rat
Alternative names: HIST1 cluster, H3J Histone gene cluster 1, H3 histone family, member E Histone gene cluster 1, H3G H3 histone family, member J HIST1 cluster, H3E HIST1 cluster, H3I Histone gene cluster 1, H3C FLJ92264 H3 histone family, member A H3 histone family, member B H3 histone family, member C H3 histone family, member D H3 histone family, member F H3 histone family, member H H3 histone family, member I H3 histone family, member K H3 histone family, member L H3 histone family, member T H3 histone, family 3A H3.1 H3.3A H3/a H3/b H3/c H3/d h3/f H3/h H3/i H3/j H3/k H3/l H3/t H31_HUMAN H3F1K H3F3 H3F3A H3FA H3FB H3FC H3FD H3FF H3FH H3FI H3FJ H3FK H3FL HIST1 cluster, H3A HIST1 cluster, H3B HIST1 cluster, H3C HIST1 cluster, H3D HIST1 cluster, H3F HIST1 cluster, H3G HIST1 cluster, H3H HIST1H3A HIST1H3B HIST1H3C HIST1H3D HIST1H3E HIST1H3F HIST1H3G HIST1H3H HIST1H3I HIST1H3J HIST......
Images
R1110-1_1.jpg Fig1: Western blot analysis of Histone H3 (mono methyl K4) 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.
R1110-1_2.png Fig2: Western blot analysis of Histone H3 (mono methyl K4) on PC-12 cell lysates with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 2 minutes;

10% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (R1110-1) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
R1110-1_3.jpg Fig3: ICC staining of Histone H3 (mono methyl K4) 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).
R1110-1_4.png Fig4: Immunocytochemistry analysis of MCF-7 cells labeling Histone H3 (mono methyl K4) with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
R1110-1_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Histone H3 (mono methyl K4) 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.
R1110-1_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Histone H3 (mono methyl K4) 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.
R1110-1_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Histone H3 (mono methyl K4) 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.
R1110-1_8.jpg Fig8: Flow cytometric analysis of Histone H3 (mono methyl K4) 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).
R1110-1_9.jpg Fig9: Dot blot analysis of Histone H3 (mono methyl K4) on different proteins with Rabbit anti-Histone H3 (mono methyl K4) antibody (R1110-1) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution for 1 hour at room temperature.

Lane 1: Unmodified Histone H3 (negative)
Lane 2: Mono-Methyl-Histone H3 (Lys4) (positive)
Lane 3: Di-Methyl-Histone H3 (Lys4) (negative)
Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative)
Lane 5: Acetyl-Histone H3 (Lys4) (negative)

Proteins loading: 100ng, 25ng, 5ng;

Blocking and dilution buffer: 5% NFDM/TBST;

Exposure time: 50 seconds.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.