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Purified anti-Histone H3.1 Phospho (Ser28) Antibody

Pricing & Availability
Clone
5D10D4 (See other available formats)
Regulatory Status
RUO
Other Names
Histone-H3
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1_5D10D4_Purified_Histone_H3point1_Phospho_Antibody_1_062416
HeLa cells were stimulated with (top) or without (bottom) nocodozole for 24 hours, then fixed and permeabilized with cold 70% ethanol, then intracellularly stained with DAPI and purified anti-Histone H3.1 Phospho (Ser28) antibody (clone 5D10D4), followed by anti-rat IgG PE.
  • 1_5D10D4_Purified_Histone_H3point1_Phospho_Antibody_1_062416
    HeLa cells were stimulated with (top) or without (bottom) nocodozole for 24 hours, then fixed and permeabilized with cold 70% ethanol, then intracellularly stained with DAPI and purified anti-Histone H3.1 Phospho (Ser28) antibody (clone 5D10D4), followed by anti-rat IgG PE.
  • 2_5D10D4_Purified_Histone_H3point1_Phospho_Antibody_2_062416
  • 3_5D10D4_Purified_Histone_H3point1_Phospho_Antibody_3_062416
    Total lysates (15 µg protein) from HeLa (lane 1) and HeLa cells treated with Nocodazole for 24 hours (lane 2) were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1 µg/mL Purified anti-Histone H3.1 Phospho (ser28) Antibody, clone 5D10D4 (upper), or 1:2000 diluted anti-GAPDH Antibody (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-Rat-IgG secondary antibody conjugated to HRP for the anti-Histone H3.1 Phospho (ser28) Antibody, or 1:3000 diluted donkey anti-Rabbit-IgG secondary antibody conjugated to HRP for anti-GAPDH Antibody. Lane M: Molecular weight ladder.
  • 4_5D10D4_GoChIP_H31_Phospho_Antibody_042617
    Chromatin Immunoprecipitation (ChIP) was performed using commercial Protein-G coated 96 well high-throughput ChIP assay kit by loading 3µg of cross-linked chromatin samples from HeLa cells treated with Nocodazole with either A) 1:50 dilution of Go-ChIP-Grade™ Purified anti-Histone H3.1 Phospho (Ser28) Antibody (Clone 5D10D4), B) equal amount of Purified Rat IgG2a, κ Isotype Control Antibody, or C) competitor’s ChIP-grade Purified anti-Histone H3.1 Phospho (Ser28) Antibody and D) equal amount of matched Isotype Control Antibody as recommended by the manufacturer. The enriched DNA was purified and quantified by real-time qPCR using primers targeting human TNFα gene region. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the 5% of total amount of input chromatin.
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687602 100 µg £238
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Description

Histone proteins are classified into core histones (H2A, H2B, H3, H4) and linker histones (H1, H5). Core histones form an octamer, which contains two H2A-H2B dimers and one H3-H4 tetramer. Core histones are predominantly globular except for the unstructured N-terminal tails. Posttranslational modifications, such as acetylation, methylation, phosphorylation, ubiquitination, SUMOylation and ADP-ribosylation occur in histone tails.

Histone modifications induce changes of chromatin structure and thereby affect the accessibility of transcription factors, nuclear proteins and enzymes to genomic DNA, resulting in gene activation or repression. It is known that histone modifications play critical roles in DNA repair, DNA replication, transcription regulation, alternative splicing and chromosome condensation and some diseases including autoimmune diseases and cancers.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Histone H3.1 S28ph peptide (21-39)(ATKAARK(phS)APATGGVKKPH), Freund's complete adjuvant.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ICFC - Quality tested
WB, ChIP - Verified
Direct ELISA, ICC - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. For ChIP applications, the suggested dilution for ChIP application is 1:50-1:100 by volume. For Western blotting, the suggested use of this reagent is 0.5 - 2.0 µg/mL. It is recommended that the reagent be titrated for optimal performance for each application.

RRID
AB_2616945 (BioLegend Cat. No. 687602)

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. Therefore, histones limit the accessibility of DNA by 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, Cell Cycle/DNA Replication, Cell Proliferation and Viability
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. Yoshimi T, et al. 2013. Monoclon. Immunodiagn. Immunother. 32:119.

Gene ID
8350 View all products for this Gene ID
UniProt
View information about Histone on UniProt.org
Go To Top Version: 7    Revision Date: 08/24/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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.
If you need assistance with selecting the best format contact our expert technical support team.

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