Purified anti-Histone H3-Phosphorylated (Ser28) Antibody

Pricing & Availability
Clone
HTA28 (See other available formats)
Regulatory Status
RUO
Other Names
Histone-H3
Isotype
Rat IgG2a, κ
HTA28_Pure_Histone_Antibody_1_031919
Whole cell extracts (15 µg protein) from untreated (-) or nocodazole-treated (+, 300 ng/mL overnight) HeLa cells were resolved on a 4-12% Bis-Tris gel, transferred to a nitrocellulose membrane and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-Histone H3 Phospho (Ser28) Antibody, clone HTA28, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ anti-β-Actin was used as a loading control at a 1:25000 dilution. Lane M: Molecular Weight marker.
  • HTA28_Pure_Histone_Antibody_1_031919
    Whole cell extracts (15 µg protein) from untreated (-) or nocodazole-treated (+, 300 ng/mL overnight) HeLa cells were resolved on a 4-12% Bis-Tris gel, transferred to a nitrocellulose membrane and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-Histone H3 Phospho (Ser28) Antibody, clone HTA28, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ anti-β-Actin was used as a loading control at a 1:25000 dilution. Lane M: Molecular Weight marker.
  • HTA28_Pure_Histone_Antibody_2_122117
    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 1:2500 diluted (0.2 µg/ml) Histone H3-Phosphorylated (Ser28) Antibody (clone HTA28, lower) overnight at 4 degrees followed by Alexa Fluor® 594 (red) conjugated goat anti-rat IgG (Cat. No. 405422) incubation for one hour at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective. The images displayed HeLa cells during cell cycle progression.
  • HTA28_Pure_Histone_Antibody_3_122117
    Untreated HeLa (Panel A) and overnight nocodazoled-treated HeLa (Panel B) were stained with purified rat monoclonal antibody against phospho-H3 (Ser28) (clone HTA28), followed by Alexa Fluor® 488 anti-alpha-tubulin, DyLight™ 594 goat anti-rat-IgG and DAPI.
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Cat # Size Price Quantity Check Availability
641001 25 µg $176.00
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641002 100 µg $411.00
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Description

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. During mitosis, H3 is phosphorylated at serine 28.This phosphorylation coincides with chromosome condensation initiated at prophase and disappears at late anaphase. H3 has been demonstrated to be phosphorylated by the action of MLTK-α (mixed linage kinase-like mitogen activated protein triple kinase α) in response to ultraviolet B light and epidermal growth factor, as well as Aurora-B during mitosis.

Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Synthetic peptide conjugated to KLH, corresponding to amino acids 23-35 of human histone H3.
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide. Final antibody concentration is 0.5 mg/ml.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
Upon receipt, store between 2°C and 8°C.
Application

WB - Quality tested
CyTOF®, ICC - Verified
IP, ICFC - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 1.0 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone is not recommended for ChIP (Chromatin Immunoprecipitation) assays (as determined by in-house testing).

Application References

(PubMed link indicates BioLegend citation)
  1. Hirata A, et al. 2004. J. Histochem. Cytochem. 52:1503.
  2. Goto H, et al. 1999. J. Biol. Chem. 274:25543.
  3. Ozawa K. 2008. Cytometry A 73:517.
  4. Goode NJ, et al. 2014. PLoS Genet. 10:1004323. PubMed
Product Citations
  1. Hutton C, et al. 2021. Cancer Cell. 39:1227. PubMed
  2. de Vargas Roditi L, et al. 2022. Cell Rep Med. 3:100604. PubMed
  3. Kuo A, et al. 2022. Elife. 11:. PubMed
  4. Rovira-Clavé X, et al. 2022. Cancer Cell. 40:1423. PubMed
  5. Fischer JR, et al. 2023. Cell Rep Med. 4:100977. PubMed
  6. Lun XK et al. 2019. Mol Cell. 74(5):1086-1102 . PubMed
  7. Damond N, et al. 2019. Cell Metab. 29:755. PubMed
  8. Baran N, et al. 2022. Nat Commun. 13:2801. PubMed
  9. Tognetti M, et al. 2021. Cell Systems. 12(5):401-418.e12. PubMed
  10. Godde N, et al. 2014. PLoS Genet. 10:1004323. PubMed
  11. Bouchard G, et al. 2022. Cancer Res. 82:648. PubMed
RRID
AB_1227660 (BioLegend Cat. No. 641001)
AB_1227660 (BioLegend Cat. No. 641002)

Antigen Details

Structure
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, 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.

Regulation
H3 is regulated by acetylation, methylation, citrullination, phosphorylation, and ubiquitination.
Gene ID
8290 View all products for this Gene ID
UniProt
View information about Histone on UniProt.org
Go To Top Version: 4    Revision Date: 03/19/2019

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This data display is provided for general comparisons between formats.
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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