H3K27me3S28p Antibody
Polyclonal antibody raised in rabbit against Histone H3 (trimethyl Lys27, p Ser28), using a KLH-conjugated synthetic peptide.
| Lot | 001 |
|---|---|
| Concentration | 0.66 μg/μl |
| Species reactivity | Human, mouse, C. elegans, rat, chicken, Xenopus, Drosophila, plant |
| Type | Polyclonal |
| Purity | Affinity purified |
| Host | Rabbit |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution * | References |
|---|---|---|
| ChIP | 2-5 μg/million cells | Fig 1 |
| Immunofluorescence | 1:200 | Fig 2, 3 |
| Western Blotting | 1:500 | Fig 4 |
| Immunochemistry | 1:50 | |
| Dot Blotting | 1:500 - 1:1,000 | Fig 5 |
Validation Data
Figure 1. ChIP
Chromatin Immunoprecipitation using the H3K27me3S28p antibody. Chromatin from one million formaldehyde cross-linked HeLa cells was used with 2 μg of H3K27me3S28p and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative PCR and normalized to the input chromatin.
Figure 2. Immunofluorescence
Immunofluorescence of H3K27me3S28p antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:200 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K27me3S28p is nuclear and chromosomal. Staining: Histone H3K27me3S28p is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).
Figure 3. Immunofluorescence
Immunofluorescence of H3K27me3S28p antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:200 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K27me3S28p is nuclear and chromosomal. Staining: Histone H3K27me3S28p is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 550 (red).
Figure 4. Western Blot
Western Blot of H3K27me3S28p antibody. Lane 1: HeLa histone extracts. Lane 2: NIH-3T3 histone extracts. Lane 3: C. elegans embryo cell lysate. Load: 30 μg per lane. Primary antibody used at 1 μg/ml overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/ Observed size: ~15 kDa. Other band(s): None.
Figure 5. Dot Blot
Dot Blot of H3K27me3S28p antibody. Lane 1: S28p/K27 unmodified. Lane 2: S28p N-Term. Lane 3: S28p C-term. Lane 4: K27Me3. Lane 5: S28p/K27Me3. Load: 1, 10, and 100 picomoles of peptide. Primary antibody at 1 μg/ml for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT.
Target Description
Chromatin is the arrangement of DNA and proteins in which chromosomes are formed. Correspondingly, chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is a very dynamic structure in which numerous post-translational modifications work together to activate or repress the availability of DNA to be copied, transcribed, or repaired. These marks decide which DNA will be open and commonly active (euchromatin) or tightly wound to prevent access and activation (heterochromatin). Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Specifically, trimethylation of K27 is associated with gene silencing, whereas pS28 is associated with mitotis and immediate early genes.
Applications
| IF Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more |
| WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more |
| DB Dot blotting Read more |
| ChIP-qPCR (ab) Read more |
Documents
| Datasheet H3K27me3S28p C15410293 DATASHEET Datasheet description | Download |
Publications
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