Histone H3 (di methyl K4) Rabbit Polyclonal Antibody
cat.: R1110-3
| 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℃. 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 of the N terminal residues of Human Di-mehtyl-Histone H3(Lys4). |
| Positive control: | Human liver tissue lysate, F9 cell lysate, MCF7, NIH/3T3, human testis tissue, mouse testis tissue, rat testis tissue. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC Dot Blot |
1:1,000 1:500-1:5,000 1:50-1:200 11,000 1:1,000 |
| Uniprot #: | SwissProt: P68431 Human | P68433 Mouse | Q6LED0 Rat |
| Alternative names: | Histone H3/a Histone H3/b Histone H3/c Histone H3/d Histone H3/f Histone H3/h Histone H3/i Histone H3/j Histone H3/k Histone H3/l H3K4me2 |
Images
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Fig1:
Western blot analysis of Histone H3 (di 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 was used at a 1:1,000 dilution 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: Lane 1: Human liver tissue lysate, untreated Lane 2: Histone lysate, purified from 293T Lane 3: F9 cell lysate, untreated Lane 4: F9 cell lysate, treated with Histone H3 peptide-unmodifed Lane 5: F9 cell lysate, treated with Histone H3 peptide-di-methyl K4 |
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Fig2:
Immunocytochemistry analysis of MCF7 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/5,000 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 (R1110-3) at 1/5,000 dilution in 1% BSA in PBST 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H3 (di methyl K4) with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/500 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 (R1110-3) at 1/500 dilution in 1% BSA in PBST 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. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/3,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-3) at 1/3,000 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/3,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-3) at 1/3,000 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) at 1/3,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (R1110-3) at 1/3,000 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. |
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Fig7:
Dot blot analysis of Histone H3 (di methyl K4) on different proteins with Rabbit anti-Histone H3 (di methyl K4) antibody (R1110-3) 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) (negative) Lane 3: Di-Methyl-Histone H3 (Lys4) (positive) 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: 1 minute 30 seconds. |
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Fig8:
Flow cytometric analysis of MCF7 cells labeling Histone H3 (di methyl K4). Cells were fixed and permeabilized. Then stained with the primary antibody (R1110-3, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig9:
Flow cytometric analysis of NIH/3T3 cells labeling Histone H3 (di methyl K4). Cells were fixed and permeabilized. Then stained with the primary antibody (R1110-3, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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