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Histone H3 dimethyl Lys27 antibody (pAb)
| Catalog No: 39245 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39246 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | Immunofluorescence |
|---|---|---|
| 1:500 - 1:1,000 | 5 - 10 µl per ChIP | 1:200 - 1:500 |
for Histone H3 dimethyl Lys27 antibody (pAb)Background
Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure. Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.
The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Lysine 27 of histone H3 can be mono-, di- or trimethylated (Histone H3 monomethyl Lys27, Histone H3 dimethyl Lys27 or Histone H3 trimethyl Lys27) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.
for Histone H3 dimethyl Lys27 antibody (pAb)Immunogen
This Histone H3 dimethyl Lys27 antibody was raised against a peptide including dimethyl-lysine 27 of histone H3.
for Histone H3 dimethyl Lys27 antibody (pAb)Application Notes
This Histone H3 dimethyl Lys27 antibody (H3K27me2) has been validated for use in Western blotting (1:500 - 1:1,000 dilution), chromatin IP (5 - 10 µl per ChIP) and immunofluorescence (1:200 - 1:500 dilution).
for Histone H3 dimethyl Lys27 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H3 dimethyl Lys27 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Catalog No. 36200) can be used as a positive control for Histone H3 dimethyl Lys27 antibody.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3 dimethyl Lys27 antibody (pAb).
Compatible Secondary Antibodies
Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H3 dimethyl Lys27 antibody (pAb).
Histone H3 dimethyl Lys27 antibody tested by Western blot.
Detection of Histone H3 dimethyl Lys27 by Western blot. The analysis was performed using Histone H3 dimethyl Lys27 antibody at a 1:1,000 dilution.
Lane 1: HeLa acid extract (20 µg per lane).
Lane 2: Recombinant histone H3 (200 ng).
Histone H3 dimethyl Lys27 antibody tested by immunofluorescence.
Staining of HeLa cells with Histone H3 dimethyl Lys27 antibody (1:500 dilution, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).
Histone H3 dimethyl Lys27 antibody tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys27 antibody for dimethyl Lys27 histone H3. Methylated peptides corresponding to the immunogen and related sequences derived from histone H3 were spotted onto PVDF and probed with the antibody at a 1:5,000 dilution. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Unmodified lysine 4 peptide.
Lane 2: Monomethyl lysine 4 peptide.
Lane 3: Dimethyl lysine 4 peptide.
Lane 4: Trimethyl lysine 4 peptide.
Lane 5: Unmodified lysine 9 peptide.
Lane 6: Monomethyl lysine 9 peptide.
Lane 7: Dimethyl lysine 9 peptide.
Lane 8: Trimethyl lysine 9 peptide.
Lane 9: Unmodified lysine 27 peptide.
Lane 10: Monomethyl lysine 27 peptide.
Lane 11: Dimethyl lysine 27 peptide.
Lane 12: Trimethyl lysine 27 peptide.
Histone H3 dimethyl Lys27 antibody tested by ChIP.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and 50 µl of Ready-to-ChIP HeLa Chromatin (Catalog No. 53015) per ChIP. Subsequent to the ChIP reaction, DNA was purified from the immunoprecipitated chromatin and a region approximately 1000 base pairs 5′ of the transcriptional start site of the HoxD13 gene was amplified by PCR.
Lane 1: ChIP using negative control rabbit IgG.
Lane 2: ChIP using 10 µl of Histone H3 dimethyl Lys27 antibody.
Lane 3: PCR input control.
for Histone H3 dimethyl Lys27 antibody (pAb)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H3 dimethyl Lys27 antibody (pAb)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
