Application
ICC/IF analysis of U2OS cells using Histone H3K4me2 (di-Methyl Lys4) antibody (red) and DAPI (blue). Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labeled with Histone H3K4me2 (di-Methyl Lys4) antibody at a dilution of 1:5,000. Additional stainings of the cells with Histone H3K4me2 (di-Methyl Lys4) antibody after incubation of the antibody with 5 ng/μl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K4 were also performed.Application
Dot blot analysis of peptides containing modifications of histone H3 and H4 and the unmodified H3K4 using H3K4me2 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 histone extracts from HeLa cells (15 μg) using H3K4me2 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.Application
ChIP was performed with HeLaS3 extracts and either control IgG (2 μg) or Histone H3K4me2 (di-Methyl Lys4) antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to the promoter and coding region of the active GAPDH gene, the promoter of the active c-fos gene, and to the coding region of the inactive TSH2B gene.| Product Name | Histone H3K4me2 (dimethyl Lys4) antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to Histone H3K4me2 (dimethyl Lys4)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 histone H3 containing the dimethylated lysine 4 (H3K4me2), using a KLH-conjugated synthetic peptide. |
| Modification | dimethyl Lys4 |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | Dot,ELISA,ICC/IF,WB,ChIP |
| ChIP/ChIP-seq (5 µg/ChIP), ELISA (1:500), Dot blotting (1:20,000), Western blotting (1:1,000) | |
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | histone cluster 1, H3a , H3/A , HIST1H3A , H3FA |
|---|---|
| Target | H3 (di-Methyl Lys4) |
Application
ICC/IF analysis of U2OS cells using Histone H3K4me2 (di-Methyl Lys4) antibody (red) and DAPI (blue). Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labeled with Histone H3K4me2 (di-Methyl Lys4) antibody at a dilution of 1:5,000. Additional stainings of the cells with Histone H3K4me2 (di-Methyl Lys4) antibody after incubation of the antibody with 5 ng/μl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K4 were also performed.
Application
Dot blot analysis of peptides containing modifications of histone H3 and H4 and the unmodified H3K4 using H3K4me2 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 histone extracts from HeLa cells (15 μg) using H3K4me2 antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.
Application
ChIP was performed with HeLaS3 extracts and either control IgG (2 μg) or Histone H3K4me2 (di-Methyl Lys4) antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to the promoter and coding region of the active GAPDH gene, the promoter of the active c-fos gene, and to the coding region of the inactive TSH2B gene.| Application Notes | ChIP/ChIP-seq (5 µg/ChIP), ELISA (1:500), Dot blotting (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. |
