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Applications Table
| Western Blot | Chromatin IP | Immunofluorescence |
|---|---|---|
| 1:500 - 1:1,000 | — | 1:1,000 - 1:5,000 |
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Background
Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.
Ser10 phosphorylation and Ser28 phosphorylation in the tail of H3 have very similar kinetics. Both phosphorylations occur early in mitosis when chromosomes begin to condense and during premature chromosome condensation induced in S-phase cells. These phosphorylated serines are excellent mitotic markers. In contrast to Ser10 phosphorylation, Ser28 phosphorylation has never been observed in interphase.
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Immunogen
This antibody was raised in rat against a peptide corresponding to amino acid residues 23-35 (phospho Ser 28) of human Histone H3.
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Application Notes
This Histone H3 phospho Ser28 Rat antibody (Clone HTA28) (H3S28Ph) has been validated for use in Western blotting (1:500 - 1:1,000 dilution) and immunofluorescence (1:1,000 - 1:5,000 dilution).
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Buffer
0.1 M Tris-glycine pH 7.4 containing 150 mM NaCl and 0.05% sodium azide.
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Positive Control
Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 phospho Ser28 Rat antibody (Clone HTA28).
Compatible Secondary Antibodies
Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H3 phospho Ser28 antibody (mAb).
Histone H3 phospho Ser28 antibody tested by immunofluorescence.
Top: HeLa cells stained with Histone H3 phospho Ser28 antibody at a 1:1,000 dilution.
Middle: Same cells stained with DAPI.
Bottom: Merge of both images.
Histone H3 phospho Ser28 antibody specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3 phospho Ser28 antibody was applied at a dilution of 1:3,000 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen. Recognition of the Histone H3 phospho Ser28 peptides by the antibody is inhibited by Lys27 monomethylation and blocked by Arg26 dimethylation and a citrulline at position 26.
Array Data File
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Storage
Antibodies in solution can be stored at -80°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H3 phospho Ser28 antibody (mAb) (Clone HTA28)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
