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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3T3p ChIP assays were performed using HeLa cells, the Diagenode antibody against H3T3p (cat. No. C15210001) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 μg of antibody per ChIP experiment was analyzed. IgG (1 μg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the TSH2B and HBB genes, used as positive controls, and for the promoters of the EIF4A2 and GAPDH genes, used as negative 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. Western blot analysis using the Diagenode monoclonal antibody directed against H3T3p Histone extracts from HeLa cells (lane 1) or HeLa cells treated with nocodazole (lane 2), as well as recombinant H3 (lane 3) were analysed by Western blot using the Diagenode monoclonal antibody against H3T3p (Cat. No. C15210001) diluted 1:10,000 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.
Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against H3T3p HeLa cells were stained with the Diagenode antibody against H3T3p (Cat. No. C15210001, red) diluted 1:1,000. Actin was stained with fluorescein phalladoin (green).
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.
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Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
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