Histone H3 [p Ser28] Antibody (HTA28)
Novus Biologicals, part of Bio-Techne | Catalog # NB600-1168
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human, Mouse, Bovine, Hamster
Cited:
Human, Mouse
Applications
Validated:
Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Microarray, Western Blot
Cited:
IF/IHC, Immunohistochemistry-Frozen
Label
Unconjugated
Antibody Source
Monoclonal Rat IgG2A Clone # HTA28
Concentration
0.5 mg/ml
Product Specifications
Immunogen
This Histone H3 [p Ser28] antibody (HTA28) was raised against synthetic peptide conjugated to KLH, corresponding to amino acids 23-35 (pSer28) of Human Histone H3.
Modification
p Ser28
Localization
Nuclear
Specificity
Histone H3 [p Ser28] antibody (HTA28) does not detect the unphosphorylated epitope. It detects the phosphorylated histone molecule at the onset of mitosis (prophase, metaphase and weaker at the beginning of anaphase), but not during late anaphase.
Clonality
Monoclonal
Host
Rat
Isotype
IgG2A
Theoretical MW
15 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for Histone H3 [p Ser28] Antibody (HTA28)
Western Blot: Histone H3 [p Ser28] Antibody (HTA28) [NB600-1168] - Whole extract of HeLa cell treated with nocodazole was separated on SDS-PAGE and probed with Monoclonal Anti-phospho-Histone H3 (pSer28), Clone: HTA28. The antibody was developed using Goat Anti-Rat IgG-Peroxidase and a chemiluminescent substrate.Lanes:1. Antibody dilution 0.5 ug/mL2. Antibody dilution 1 ug/mL3. Antibody dilution 2 ug/mL
Immunocytochemistry/Immunofluorescence: Histone H3 [p Ser28] Antibody (HTA28) [NB600-1168] - HeLa cells were fixed and permeabilized with methanol followed by methanol:acetone. Fixed cells were stained with 5 ug/mL Monoclonal Anti-phospho-Histone H3 (pSer28), Clone: HTA28. The antibody was developed using Goat Anti-Rat IgG, FITC conjugate (green). Cells were counterstained with DAPI (blue) to stain nuclei.
Flow Cytometry: Histone H3 [p Ser28] Antibody (HTA28) [NB600-1168] - FACS profile of human leukemic cells with Anti-phospho-Histone H3.
Applications for Histone H3 [p Ser28] Antibody (HTA28)
Application
Recommended Usage
Flow Cytometry
1:10-1:1000
Immunocytochemistry/ Immunofluorescence
1:10-1:2000
Western Blot
0.5-1.0 ug/ml
Application Notes
For ICC: use 3.7% formaldehyde-methanol fixation Use in Immunohistochemistry reported in scientific literature (PMID 25258086)
Formulation, Preparation, and Storage
Purification
Protein A or G purified
Formulation
10mM PBS (pH 7.4) and 1.0% BSA
Preservative
0.9% Sodium Azide
Concentration
0.5 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: Histone H3
Histones are nuclear proteins responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Changes in chromatin structure play a large role in the regulation of transcription. The chromatin fibers are compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.
Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Posttranslational modifications such as acetylation of core histones regulates gene expression, thus altering protein function and regulation (1). Histone H3 is primarily acetylated at lysines 9, 14, 18, and 23 and have a theoretical molecular weight of 15 kDa. Acetylation at lysine 9 and 14 appears to control histone deposition, chromatin assembly and active transcription. Methylation of arginine residues within histone H3 has also been linked to transcription regulation. Histone H3 has been linked to various types of cancer as a biomarker through the aberrant expression of histone deacetylase (HDAC) enzymes and changes to chromatins (2-4).
References
1. Zhang, Y. X., Akumuo, R. C., Espana, R. A., Yan, C. X., Gao, W. J., & Li, Y. C. (2018). The histone demethylase KDM6B in the medial prefrontal cortex epigenetically regulates cocaine reward memory. Neuropharmacology, 141, 113-125. doi:10.1016/j.neuropharm.2018.08.030
2. Nandakumar, V., Hansen, N., Glenn, H. L., Han, J. H., Helland, S., Hernandez, K, ...Meldrum, D. R. (2016). Vorinostat differentially alters 3D nuclear structure of cancer and non-cancerous esophageal cells. Sci Rep, 6, 30593. doi:10.1038/srep30593
3. Zhou, M., Li, Y., Lin, S., Chen, Y., Qian, Y., Zhao, Z., & Fan, H. (2019). H3K9me3, H3K36me3, and H4K20me3 Expression Correlates with Patient Outcome in Esophageal Squamous Cell Carcinoma as Epigenetic Markers. Dig Dis Sci, 64(8), 2147-2157. doi:10.1007/s10620-019-05529-2
4. Li, Y., Guo, D., Sun, R., Chen, P., Qian, Q., & Fan, H. (2019). Methylation Patterns of Lys9 and Lys27 on Histone H3 Correlate with Patient Outcome in Gastric Cancer. Dig Dis Sci, 64(2), 439-446. doi:10.1007/s10620-018-5341-8
Alternate Names
H3F3A, H3S28p
Gene Symbol
H3-3A
UniProt
Additional Histone H3 Products
Product Documents for Histone H3 [p Ser28] Antibody (HTA28)
Product Specific Notices for Histone H3 [p Ser28] Antibody (HTA28)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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