WB
Western blot analysis of Histone H3 (mono methyl K27) on different lysates with Rabbit anti-Histone H3 (mono methyl K27) antibody at 1/1,000 dilution. Lane 1: MCF7 cell lysate, Lane 2: SH-SY5Y cell lysate, Lane 3: HeLa cell lysate, Lane 4: A549 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 50 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 2% BSA/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H3 (mono methyl K27) with Rabbit anti-Histone H3 (mono methyl K27) antibody 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 (mono methyl K27) antibody at 1/500 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.ICC/IF
Immunocytochemistry analysis of C6 cells labeling Histone H3 (mono methyl K27) with Rabbit anti-Histone H3 (mono methyl K27) antibody at 1/250 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 (mono methyl K27) antibody at 1/250 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.FC
Flow cytometric analysis of MCF7 cells labeling Histone H3 (mono methyl K27). Cells were fixed and permeabilized. Then stained with the primary antibody (1/500) (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells with Histone H3 (mono methyl K27) / Competitor C / 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 (mono methyl K27) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human Histone H3 (mono methyl K27) aa 1-50. |
| Modification | mono methyl K27 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPFCICC/IFWB |
| WB:1:1000 ICC/IF:1:100-1:500 FC:1:500 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 E pseudogene antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3F3 antibody H3FA antibody Hist1h3a antibody HIST1H3B antibody HIST1H3C antibody HIST1H3D antibody H3 histone family member E pseudogene antibody H3 histone family member A antibody H3/A antibody H31_HUMAN antibody H3F3 antibody H3FA 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 cluster 1 H3a antibody Histone H3 3 pseudogene 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 H3K27me |
|---|---|
| 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. |
WB
Western blot analysis of Histone H3 (mono methyl K27) on different lysates with Rabbit anti-Histone H3 (mono methyl K27) antibody at 1/1,000 dilution. Lane 1: MCF7 cell lysate, Lane 2: SH-SY5Y cell lysate, Lane 3: HeLa cell lysate, Lane 4: A549 cell lysate, Lane 5: NIH/3T3 cell lysate, Lane 6: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 50 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 2% BSA/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Histone H3 (mono methyl K27) with Rabbit anti-Histone H3 (mono methyl K27) antibody 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 (mono methyl K27) antibody at 1/500 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.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling Histone H3 (mono methyl K27) with Rabbit anti-Histone H3 (mono methyl K27) antibody at 1/250 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 (mono methyl K27) antibody at 1/250 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.
FC
Flow cytometric analysis of MCF7 cells labeling Histone H3 (mono methyl K27). Cells were fixed and permeabilized. Then stained with the primary antibody (1/500) (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ChIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells with Histone H3 (mono methyl K27) / Competitor C / 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 ICC/IF:1:100-1:500 FC:1:500 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
