WB
Western blot analysis of Histone H3 (di methyl K4) on different lysates with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: MCF7 cell lysate, Lane 3: HCT 116 cell lysate, Lane 4: COS-1 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 14 seconds; 4-20% 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 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour 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 rat colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour 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 NIH/3T3 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution in 1% BSA in PBST 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. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.DB
Dot blot analysis of Histone H3 (di methyl K4) on different proteins with Rabbit anti-Histone H3 (di 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) (negative), Lane 3: Di-Methyl-Histone H3 (Lys4) (positive), Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative), Lane 5: Acetyl-Histone H3 (Lys4) (negative), Lane 6: Acetyl-Histone H3 (Lys9) (negative), Proteins loading: 100ng, 25ng, 5ng, 1ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 15 seconds.ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (di methyl K4) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | Histone H3 (di methyl K4) Rabbit Polyclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide of the N terminal residues of Human Di-mehtyl-Histone H3(Lys4). |
| Modification | di methyl K4 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPFCICC/IFIHCWB |
| WB:1:1000 IHC:10000 ICC/IF:1:500-1:2000 DB:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody HIST1H3E antibody HIST1H3F antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody HIST1H3E antibody HIST1H3F antibody HIST1H3G antibody HIST1H3H antibody HIST1H3I antibody HIST1H3J antibody histone 1 H3a antibody Histone cluster 1 H3a antibody Histone H3.1 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 H3K4me2 antibody |
|---|---|
| Molecular Weight(MW) | 15kDa |
| Function | In eukaryotes, DNA is wrapped around histone octamers to form the basic unit of chromatin structure. The octamer is composed of histones H2A, H2B, H3 and H4, and it associates with approximately 200 base pairs of DNA to form the nucleosome. The association of DNA with histones results in dense packing of chromatin, which restricts proteins involved in gene transcription from binding to DNA. p300 preferentially acetylates Histone H3 at lysines 14 and 18 and Histone H4 at lysines 5 and 8. PCAF in its native form, primarily acetylates Histone H3 at lysine 14 to a monoacetylated form, and less efficiently acetylates Histone H4 at lysine 8. Histone H4 may also be acetylated at lysines 12 and 16, and the involvement of acetylated H4 with Histones H2A, H2B and H3 suggests that acetylated histones may be involved in dynamic chromatin remodeling. |
| Cellular Localization | Nucleus, Chromosome. |
| SwissProt ID | P68431 |
|---|
WB
Western blot analysis of Histone H3 (di methyl K4) on different lysates with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate, Lane 2: MCF7 cell lysate, Lane 3: HCT 116 cell lysate, Lane 4: COS-1 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: C6 cell lysate, Lysates/proteins at 10 µg/Lane. Exposure time: 14 seconds; 4-20% 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 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour 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 rat colon tissue with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour 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 NIH/3T3 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 (di methyl K4) antibody at 1/1,000 dilution in 1% BSA in PBST 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. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
DB
Dot blot analysis of Histone H3 (di methyl K4) on different proteins with Rabbit anti-Histone H3 (di 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) (negative), Lane 3: Di-Methyl-Histone H3 (Lys4) (positive), Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative), Lane 5: Acetyl-Histone H3 (Lys4) (negative), Lane 6: Acetyl-Histone H3 (Lys9) (negative), Proteins loading: 100ng, 25ng, 5ng, 1ng; Blocking and dilution buffer: 5% NFDM/TBST; Exposure time: 15 seconds.
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H3 (di methyl K4) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:1000 IHC:10000 ICC/IF:1:500-1:2000 DB:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
