Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H3K27me3 (tri-Methy Lys27) antibody (0.1, 0.5, and 1 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls MYT1 and TSH2B genes, and to negative controls GAPDH and EIF4A2 promoters.Application
ELISA was performed using a serial dilution of Histone H3K27me3 (tri-Methy Lys27) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:22,400.Application
Dot blot analysis of peptides containing modifications or unmodified sequences of histone H3 and H4 using H3K27me3 antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K27me3 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.Application
ICC/IF analysis of NIH3T3 cells using H3K27me3 antibody (green) and DAPI (blue). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H3K27me3 antibody at a dilution of 1:500.| Product Name | Histone H3K27me3 (trimethy Lys27) antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to Histone H3K27me3 (trimethy Lys27)Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq, Jul 2008] |
| Immunogen | Polyclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 27 (H3K27me3), using a KLH-conjugated synthetic peptide. |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | Dot,ELISA,ICC/IF,WB,ChIP |
| ChIP (1 µg/IP), ELISA (1:1,000), Dot blotting/Peptide array (1:20,000), Western blotting (1:1,000) | |
| Species Reactivity | Human,Mouse |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | histone cluster 1, H3a , H3/A , HIST1H3A , H3FA |
|---|---|
| Target | H3 (tri-Methy Lys27) |
Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H3K27me3 (tri-Methy Lys27) antibody (0.1, 0.5, and 1 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls MYT1 and TSH2B genes, and to negative controls GAPDH and EIF4A2 promoters.
Application
ELISA was performed using a serial dilution of Histone H3K27me3 (tri-Methy Lys27) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:22,400.
Application
Dot blot analysis of peptides containing modifications or unmodified sequences of histone H3 and H4 using H3K27me3 antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.
Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K27me3 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.
Application
ICC/IF analysis of NIH3T3 cells using H3K27me3 antibody (green) and DAPI (blue). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H3K27me3 antibody at a dilution of 1:500.| Application Notes | ChIP (1 µg/IP), ELISA (1:1,000), Dot blotting/Peptide array (1:20,000), Western blotting (1:1,000) |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide and 0.05% ProClin 300. |
