WB
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 (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody 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.WB
Western blot analysis of Histone H3 (mono methyl K4) on PC-12 cell lysates with Rabbit anti-Histone H3 (mono methyl K4) antibody at 1/500 dilution. Lysates/proteins at 10 µg/Lane. 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 at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.IHC
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 (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.IHC
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 (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.IHC
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 (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.ICC/IF
Immunocytochemistry analysis of MCF-7 cells labeling Histone H3 (mono methyl K4) with Rabbit anti-Histone H3 (mono methyl K4) antibody 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 at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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.FC
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 (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).DB
Dot blot analysis of Histone H3 (mono methyl K4) on different proteins with Rabbit anti-Histone H3 (mono methyl K4) antibody at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody 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.| Product Name | Histone H3 (mono methyl K4) Rabbit Polyclonal Antibody |
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| Antibody Type | Primary Antibodies |
| Product description | 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. |
| Immunogen | Synthetic peptide within human Histone H3 aa 1-49/136. |
| Modification | mono methyl K4 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | DBFCICC/IFIHCWB |
| WB:1:500-1:2000 IHC:1:50-1:200 ICC/IF:1:50-1:200 FC:1:50-1:100 DB:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Alternative Names | 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 |
|---|---|
| Molecular Weight(MW) | 15 kDa |
| Cellular Localization | Nucleus |
| SwissProt ID | P68431 |
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WB
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 (1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody 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.
WB
Western blot analysis of Histone H3 (mono methyl K4) on PC-12 cell lysates with Rabbit anti-Histone H3 (mono methyl K4) antibody at 1/500 dilution. Lysates/proteins at 10 µg/Lane. 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 at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.
IHC
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 (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.
IHC
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 (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.
IHC
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 (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.
ICC/IF
Immunocytochemistry analysis of MCF-7 cells labeling Histone H3 (mono methyl K4) with Rabbit anti-Histone H3 (mono methyl K4) antibody 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 at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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.
FC
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 (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).
DB
Dot blot analysis of Histone H3 (mono methyl K4) on different proteins with Rabbit anti-Histone H3 (mono methyl K4) antibody at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody 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.| Application Notes | WB:1:500-1:2000 IHC:1:50-1:200 ICC/IF:1:50-1:200 FC:1:50-1:100 DB:1:1000 |
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| Form | Liquid |
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| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
