HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with (A) Alexa Fluor® 594 Mouse IgG2b, κ Isotype Ctrl Antibody (Negative, Cat. No. 400362) or (B-D) Alexa Fluor® 594 Histone H3 Phospho (Ser10) Antibody (Clone 11D8) overnight at 4°C. Nuclei were counterstained with DAPI (Blue, Cat. No. 422801). The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (Seconds) for (A-D) is 1/400. Concentrations for (A-B) is 2 µg/ml (1:250 dilution), (C) is 0.5 µg/ml (1:1000 dilution) and (D) is 0.1 µg/ml (1:5000 dilution).
HeLa cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with (A) Alexa Fluor® 594 Mouse IgG2b, κ Isotype Ctrl Antibody (Negative, Cat. No. 400362) or (B-D) Alexa Fluor® 594 Histone H3 Phospho (Ser10) Antibody (Clone 11D8) overnight at 4°C. Nuclei were counterstained with DAPI (Blue, Cat. No. 422801). The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (Seconds) for (A-D) is 1/400. Concentrations for (A-B) is 2 µg/ml (1:250 dilution), (C) is 0.5 µg/ml (1:1000 dilution) and (D) is 0.1 µg/ml (1:5000 dilution).
Histone H3 is phosphorylated at serine 10 during mitosis and is found to be involved in transcriptional activation, chromatin decondensation, and chromosome compaction during cell division, by the action of Aurora kinase and NIMA kinases.
Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 2 - 5 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
H3 is part of the nucleosome, comprised of an octameric complex with H2A, H2B, and H4 proteins.
Distribution
Nucleus
Function
H3 is a core component of the nucleosome that serves to wrap and compact DNA into chromatin. Histones, therefore, limit the accessibility of DNA, providing mechanisms for transcription regulation, DNA repair and replication, and chromosomal stability.
Interaction
Two molecules of H3 form a heterotetramer with two molecules of H4.
Biology Area
Cell Biology, Chromatin Remodeling/Epigenetics, DNA Repair/Replication, Transcription Factors
Molecular Family
Phospho-Proteins
Antigen References
1. Choi HS, et al. 2005. J. Biol. Chem. 280:13545.
2. Goto H, et al. 2002. Genes Cells 7:11.
3. Garcia BA, et al. 2005. Biochemistry 44:13202.
4. Hans F, et al. 2001. Oncogene 20:3021.
Regulation
H3 is regulated by acetylation, methylation, citrullination, phosphorylation, and ubiquitination.
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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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