Application
ChIP was performed with HeLa nuclear extract and either 1 μl of H3K27me3S28p antibody or 5 μg of control rabbit IgG. The precipitated DNA was detected by QPCR with primer set targeting to RPL30 promoter or MyoD.Application
Dot blot analysis of peptides containing modified histone H3 and H4 and unmodified sequences of histone H3 using H3K27me3S28ph antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the peptides were spotted on a membrane for analysis.Application
ICC/IF analysis of HeLa asynchronous cells using H3K27me3S28ph antibody and DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. Cells were immunofluorescently labelled with the H3K27me3S28ph antibody diluted 1:200 (green). The nuclei were stained with DAPI (blue).Application
HeLa cells were treated with colcemid to block the cell cycle in metaphase and were fixed with formaldehyde. Chromatin from 10,000 cells was sheared and used for immunoprecipitation (IP). IP was performed with 5 μl of H3K27me3S28ph antibody. The immunoprecipitated proteins were analysed by WB with the antibody diluted 1:500 in TBS-Tween containing 5% skimmed milk. Lane 1 shows the result of the IP; a negative IP control (no antibody added) and a positive control (sheared chromatin from 10,000 cells) are shown in lane 2 and 3, respectively.Application
WB analysis of histone extracts (15 μg) from HeLa cells using H3K27me3S28ph antibody diluted 1:250 in TBS-Tween containing 5% skimmed milk. Lane 2 shows the result of the WB analysis with the antibody; lane 1 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.Application
ELISA was performed using a serial dilution of Histone H3K27me3S28ph (tri-Methyl Lys27, phospho Ser28) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.| Product Name | Histone H3K27me3S28ph (trimethyl Lys27, phospho Ser28) antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to Histone H3K27me3S28ph (trimethyl Lys27, phospho Ser28) |
| Modification | trimethyl Lys27, phospho Ser28 |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | DotELISAICC/IFIPWBChIP |
| ChIP (1 µl/ChIP), ELISA (1:100 – 1:500), Dot blotting (1:20,000), Western blotting (1:250 - 1:500), Immunofluorescence (1:200), Immunoprecipitation (5 µl/IP): |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | tri Methyl Histone H3 K27 Histone H3 phospho S28 tri-Methyl K27 phospho S28 tri-Methyl K27 Histone H3 histone cluster 1 H3a H3/A HIST1H3A phospho Histone H3 (pS28) triMethyl Lys27 Histone H3 Histone H3 tri Methyl Histone H3 (pS28) Histone H3 t |
|---|---|
| Target | Histone H3 (tri-Methyl Lys27, phospho Ser28) |
Application
ChIP was performed with HeLa nuclear extract and either 1 μl of H3K27me3S28p antibody or 5 μg of control rabbit IgG. The precipitated DNA was detected by QPCR with primer set targeting to RPL30 promoter or MyoD.
Application
Dot blot analysis of peptides containing modified histone H3 and H4 and unmodified sequences of histone H3 using H3K27me3S28ph antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the peptides were spotted on a membrane for analysis.
Application
ICC/IF analysis of HeLa asynchronous cells using H3K27me3S28ph antibody and DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. Cells were immunofluorescently labelled with the H3K27me3S28ph antibody diluted 1:200 (green). The nuclei were stained with DAPI (blue).
Application
HeLa cells were treated with colcemid to block the cell cycle in metaphase and were fixed with formaldehyde. Chromatin from 10,000 cells was sheared and used for immunoprecipitation (IP). IP was performed with 5 μl of H3K27me3S28ph antibody. The immunoprecipitated proteins were analysed by WB with the antibody diluted 1:500 in TBS-Tween containing 5% skimmed milk. Lane 1 shows the result of the IP; a negative IP control (no antibody added) and a positive control (sheared chromatin from 10,000 cells) are shown in lane 2 and 3, respectively.
Application
WB analysis of histone extracts (15 μg) from HeLa cells using H3K27me3S28ph antibody diluted 1:250 in TBS-Tween containing 5% skimmed milk. Lane 2 shows the result of the WB analysis with the antibody; lane 1 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.
Application
ELISA was performed using a serial dilution of Histone H3K27me3S28ph (tri-Methyl Lys27, phospho Ser28) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.| Application Notes | ChIP (1 µl/ChIP), ELISA (1:100 – 1:500), Dot blotting (1:20,000), Western blotting (1:250 - 1:500), Immunofluorescence (1:200), Immunoprecipitation (5 µl/IP): |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | Whole antiserum from rabbit containing 0.05% azide. |
