Rabbit Polyclonal H3 phospho S28 antibody. Suitable for WB, PepArr, ICC/IF and reacts with Human, Synthetic peptide - Human samples. Cited in 17 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
WB | PepArr | ICC/IF | |
---|---|---|---|
Human | Tested | Expected | Tested |
Drosophila melanogaster | Predicted | Predicted | Predicted |
Synthetic peptide - Human | Not recommended | Tested | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info 0.00200-0.02 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/5000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
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Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
H3FA, HIST1H3A, H3C2, H3FL, HIST1H3B, H3C3, H3FC HIST1H3C, H3C4, H3FB, HIST1H3D, H3C6, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3C10, H3FK, HIST1H3H, H3C11, H3FF, HIST1H3I, H3C12, H3FJ, HIST1H3J, H3C10, HIST1H3J, H3FJ, H3C12, HIST1H3I, H3FF, H3C6, HIST1H3A, H3FA, H3C1, HIST1H3H, H3C11, H3C2, H3FL, HIST1H3B, H3C3, H3FC, HIST1H3C, H3C4, H3FB, HIST1H3D, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3FK, Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/i, Histone H3/j, Histone H3/k, Histone H3/l, H3S28p
Rabbit Polyclonal H3 phospho S28 antibody. Suitable for WB, PepArr, ICC/IF and reacts with Human, Synthetic peptide - Human samples. Cited in 17 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
This antibody is specific for Histone H3 phosphorylated at residue Ser 28 and does not recognise the unmodified residue or another phosphorylated residue (Ser 10) on the same histone.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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Full details and terms and conditions can be found here:
Terms & Conditions.
0-4hr old wee shRNA embryos (lane 2) should display elevated phH3Ser28 levels relative to 0-4hr old EGFP shRNA embryos (lane 1). Blocked with 10% BSA.
All lanes: Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169) at 1/1000 dilution
Lane 1: Wild type 0-4 hour old fruit fly embryos.
Lane 2: 0-4 hour old fruit fly embryos expressing wee RNAi
All lanes: Donkey anti-rabbit IgG, Horseradish Peroxidase-L at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Exposure time: 10min
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab5169 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.
All lanes: Western blot - Anti-Histone H3 (phospho S28) antibody (ab5169) at 1 µg/mL
All lanes: Hela Whole Cell Lysate - Colcemid Treated at 2.5 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15 kDa
Observed band size: 17 kDa
Exposure time: 30s
All batches of ab5169 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - phospho S28 peptide (ab5499), indicating that this antibody specifically recognises the Histone H3 - phospho S28 modification.
ab17163 - Histone H3 unmodified
ab5499 - Histone H3 - phospho S28
Human Histone H3 (phospho S10) peptide ab11477 - Histone H3 - phospho S10
Indian Muntjac (top panel) and HeLa cells (bottom panel) immunofluorescently labelled with ab5169 (green) at a working dilution of 1/5000. The DNA is counterstained with DAPI and is shown in blue in the top panel and red in the bottom panel. This antibody gives a characteristic staining pattern for Histone H3 (phospho S28) whereby the signal increases in intensity during late G2 and continues to increase until metaphase. Upon entry into anaphase the signal begins to decrease until reaching basal levels by early G1. 100x magnification.
In situ peptide competition was perfomed on paraformaldehyde-fixed HeLa cells. Four 25μl aliquots were made, to which 7.5μg (1.5μl) of no peptide, H3 unmodified peptide (Human Histone H3 (unmodified ) peptide ab2623), H3 phospho S10 peptide (Human Histone H3 (phospho S10) peptide ab11477) or H3 phospho S28 peptide (ab5499) was added, mixed by vortexing and incubated for 1 hour at room temperature. Cells on glass coverslips were fixed with 4% paraformaldehyde (10min) and gently washed twice with PBS, then permeabilized with 0.5% Triton X-100 in PBS (10min) and gently washed three times with PBS. The cells were immunofluorescently labeled with either the peptide-competed antibody or the control antibody (i.e. no peptide) for 30min at room temperature, washed briefly with PBS containing 0.1% Triton X-100 (1 min) and twice with PBS. The cells were then incubated with an appropriate dilution of a secondary antibody at room temperature for 30min, rinsed as above and mounted using a 90% glycerol in PBS mount
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