H3pan Antibody
This antibody has been raised in rabbit against two KLH-conjugated synthetic peptides containing an unmodified sequence from the central part and from the C-terminus of histone H3, respectively.
| Lot | A2566-001 |
|---|---|
| Concentration | Not determined |
| Species reactivity | Human, zebrafish, Daphnia: positive. Other species: not tested. |
| Type | Polyclonal |
| Purity | Whole antiserum |
| Host | Rabbit |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP | 1 μl/ChIP | Fig 1 |
| ELISA | 1:10,000 | Fig 2 |
| Western Blotting | 1:500 | Fig 3 |
* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.
Validation Data
Figure 1. ChIP results obtained with the Diagenode antibody directed against H3pan
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (Cat. No. C15310135) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μl per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3pan (Cat. No. C15310135). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.
Figure 3. Western blot analysis using the Diagenode antibody directed against H3pan
Whole cell extracts from HeLa cells (25 μg) were analysed by Western blot using the Diagenode antibody against H3pan (Cat. No. C15310135) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Target Description
Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.
Applications
| ELISA Enzyme-linked immunosorbent assay. 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 |
| ChIP-qPCR (ab) Read more |
Documents
| Datasheet H3pan C15310135 DATASHEET Datasheet description | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
Safety sheets
Publications
How to properly cite our product/service in your workWe strongly recommend using this: H3pan Antibody (Hologic Diagenode Cat# C15310135 Lot# A2566-001). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
Epigenetic, transcriptional and phenotypic responses in Daphnia magna exposed to low-level ionizing radiation |
The 20S proteasome activator PA28γ controls the compaction of chromatin |
The TGF-β profibrotic cascade targets ecto-5'-nucleotidase gene in proximal tubule epithelial cells and is a traceable marker of progressive diabetic kidney disease. |
Inhibition of methyltransferase activity of enhancer of zeste 2 leads to enhanced lipid accumulation and altered chromatin status in zebrafish. |
lncRNA KHPS1 Activates a Poised Enhancer by Triplex-Dependent Recruitment of Epigenomic Regulators. |
Gamma radiation induces locus specific changes to histone modification enrichment in zebrafish and Atlantic salmon. |
SIRT7-Dependent Deacetylation of Fibrillarin Controls Histone H2A Methylation and rRNA Synthesis during the Cell Cycle. |
HIV-2/SIV viral protein X counteracts HUSH repressor complex. |
The transcription factors Runx3 and ThPOK cross-regulate acquisition of cytotoxic function by human Th1 lymphocytes |
The transcription factors Runx3 and ThPOK cross-regulate acquisition of cytotoxic function by human Th1 lymphocytes. |
Phenotypic Plasticity through Transcriptional Regulation of the Evolutionary Hotspot Gene tan in Drosophila melanogaster |
Heat shock represses rRNA synthesis by inactivation of TIF-IA and lncRNA-dependent changes in nucleosome positioning |
Role of Annexin gene and its regulation during zebrafish caudal fin regeneration |
Standardizing chromatin research: a simple and universal method for ChIP-seq |
Nurr1 and Retinoid X Receptor Ligands Stimulate Ret Signaling in Dopamine Neurons and Can Alleviate α-Synuclein Disrupted Gene Expression |
The Integrase Cofactor LEDGF/p75 Associates with Iws1 and Spt6 for Postintegration Silencing of HIV-1 Gene Expression in Latently Infected Cells. |
Endonuclease G preferentially cleaves 5-hydroxymethylcytosine-modified DNA creating a substrate for recombination. |
Nitric oxide-induced neuronal to glial lineage fate-change depends on NRSF/REST function in neural progenitor cells. |