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Histone H3 trimethyl Lys27 antibody (mAb)
| Clone: MABI 0323 |
| Catalog No: 61017 | Format: 100 µg | $340 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | IF | IP |
|---|---|---|---|
| 0.5 - 2 µg/ml | 2 - 10 µg per ChIP | 0.5 - 1 µg/ml | 1 - 5 µg per IP |
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)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 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)Immunogen
This Histone H3 trimethyl Lys27 antibody was raised against a peptide containing trimethyl Lys27 of human Histone H3.
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)Application Notes
This Histone H3 trimethyl Lys27 antibody chromatin IP (ChIP) (2 - 10 µg per ChIP), immunofluorescence (0.5 - 1 µg/ml), immunoprecipitation (1 - 5 µg per IP) and Western blotting (0.5 - 2 µg/ml). See references for more details.
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)Buffer
PBS pH 7.5, 30% glycerol, 0.035% sodium azide
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)Positive Control
Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 trimethyl 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 trimethyl Lys27 antibody (mAb).
Bridging Antibody to Improve ChIP, MeDIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3 trimethyl Lys27 antibody (mAb).
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 trimethyl Lys27 antibody (mAb).
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)References
The creation of this antibody was described in Kimura, H. et al (2008) Cell Struct Func 33: 61-73.
This antibody has been used in the following references:
ChIP: Ohhata T et al (2008) Development 135: 227-235; Luco RF et al (2010) Cell 327: 996-1000.
IF: Hayashi-Takanaka Y et al (2011) Nucl Acid Res 39: 6475-6488.
WB: Hayashi-Takanaka Y et al (2011) Nucl Acid Res 39: 6475-6488.
Western blot of Histone H3 trimethyl Lys27 antibody.
HeLa nuclear extract (20 µg per lane) probed with Histone H3 trimethyl Lys27 antibody (2 µg/ml dilution).
Dot blot of Histone H3 trimethyl Lys27 antibody.
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys27 antibody for trimethyl Lys27 of histone H3. Recombinant methylated histone proteins corresponding to the immunogen and related sequences were spotted onto PVDF and probed with Histone H3 trimethyl Lys27 at 2 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row. Lane 1: unmodified H3 protein. Lane 2: monomethyl Lys4 protein. Lane 3: dimethyl Lys4 protein. Lane 4: trimethyl Lys4 protein. Lane 5: monomethyl Lys9 protein. Lane 6: dimethyl Lys9 protein. Lane 7: trimethyl Lys9 protein. Lane 8: monomethyl Lys27 protein. Lane 9: dimethyl Lys27 protein. Lane 10: trimethyl Lys27 protein.
for Histone H3 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)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 trimethyl Lys27 antibody (Clone mAb) (Clone MABI 0323)Guarantee
This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.
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
