- Clone
- 5D10D4 (See other available formats)
- Regulatory Status
- RUO
- Other Names
- Histone-H3
- Isotype
- Rat IgG2a, κ
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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. -
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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. -
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.
| Cat # | Size | Price | Quantity Check Availability | ||
|---|---|---|---|---|---|
| 687602 | 100 µg | $347.00 | |||
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
- 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
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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
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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
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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
Other Formats
View All Histone Reagents Request Custom Conjugation| Description | Clone | Applications |
|---|---|---|
| Purified anti-Histone H3.1 Phospho (Ser28) | 5D10D4 | ICFC,WB,ChIP,Direct ELISA,ICC |
| Alexa Fluor® 647 anti-Histone H3.1 Phospho (Ser28) | 5D10D4 | ICFC |
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Purified anti-Histone H3.1 Phospho (Ser28)
HeLa cells were stimulated with (top) or without (bottom) no... 
Total lysates (15 µg protein) from HeLa (lane 1) and HeLa ce... 
Chromatin Immunoprecipitation (ChIP) was performed using com... -
Alexa Fluor® 647 anti-Histone H3.1 Phospho (Ser28)
HeLa cells were stimulated with (top) or without (bottom) no... 