Anti h3s10ph

Manufactured by Merck Group

Sourced in United States

Anti-H3S10ph is a research-use only antibody targeting phosphorylation of histone H3 at serine 10. It is designed for use in various laboratory techniques to detect and analyze this specific histone modification.

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Lab products found in correlation

Benzonase nuclease, by Merck Group (2 mentions) Clarity ecl, by Bio-Rad (2 mentions) Anti mouse igg hrp, by Cell Signaling Technology (2 mentions) Nupage 4 12 bis tris protein gel, by Thermo Fisher Scientific (2 mentions) Anti rabbit igg hrp, by Cell Signaling Technology (2 mentions) Ab21686, by Abcam (2 mentions) B 5 1 2, by Merck Group (2 mentions) Anti vrk2, by Santa Cruz Biotechnology (2 mentions) Anti vinculin, by Cell Signaling Technology (2 mentions) Anti mpm2, by Merck Group (2 mentions) Anti acetylated α tubulin, by Merck Group (1 mentions) Monastrol, by Merck Group (1 mentions) Anti bubr1, by Abcam (1 mentions) Anti mad2, by Santa Cruz Biotechnology (1 mentions) Anti α tubulin, by Merck Group (1 mentions) Anti h3k9ac, by Merck Group (1 mentions) Anti h3k4me2, by Merck Group (1 mentions) Anti h3ac, by Merck Group (1 mentions) Anti γh2ax ab2893, by Abcam (1 mentions)

Spelling variants of this product

H3S10ph (2 citations) Rabbit anti-H3S10ph (2 citations)

5 protocols using anti h3s10ph

Immunofluorescence Analysis of Oocyte Meiosis

Cited 1 time

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Oocytes at specific stages were fixed in 4% paraformaldehyde for 10 min and permeabilized in phosphate buffered saline (PBS) with 0.1% Triton X-100 for 15 min. Oocytes were then incubated with primary antibodies followed by Alexa Fluor-conjugated 488 and 594 secondary antibodies (Jackson ImmunoResearch). DAPI was used for DNA counterstaining. At least 20 oocytes were examined for each group, unless otherwise stated. The antibodies used in this study were: anti-Zwint-1 (Abcam; 1:50), anti-BubR1 (Abcam; 1:50), anti-Mad2 (Santa Cruz; 1:50), anti-Aurora C kinase (Origene; 1:50), anti-H3S10ph (Millipore; 1:50), anti-acetylated α-tubulin (Sigma; 1:500) and anti-centromere (Antibodies Incorporated; 1:50).
For aneuploidy analysis, oocytes that had been pretreated for 2 hours in 200 μM monastrol (Sigma) were fixed and stained with the indicated antibodies, as described previously45 (link).

Woo Seo D., Yeop You S., Chung W.J., Cho D.H., Kim J.S, & Su Oh J. (2015). Zwint-1 is required for spindle assembly checkpoint function and kinetochore-microtubule attachment during oocyte meiosis. Scientific Reports, 5, 15431.

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Western Blot Analysis of Mitotic Regulators

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Cells were washed once in PBS and then lysed in 141 mM Tris Base, 106 mM Tris HCl, 2% LDS, 1% Benzonase nuclease (E1014, Millipore), pH 8.5, at room temperature for 15 min and snap frozen in liquid nitrogen. The protein concentration was determined using Rapid Gold BCA Protein Assay Kit (Pierce). Cell lysates (20 µg) were run on NuPAGE 4–12% Bis–Tris protein gels (Invitrogen) using standard procedures, transferred to 0.2 µm polyvinylidene fluoride membranes and blocked in 5% milk in TBST (0.05% Tween 20 in Tris-buffered saline) for 1 h at room temperature before overnight incubation with primary antibodies. Membranes were washed thrice in TBST then incubated with a HRP conjugated secondary antibody for 1 h at room temperature in TBST with 5% milk, and signals were detected by chemiluminescence (Clarity ECL, Biorad). Antibodies used in immunoblotting were as follows: rabbit polyclonal anti-Vinculin (Cell Signaling Technology, #13901), anti-Haspin (Abcam, ab21686, lot 156753), and anti-H3T3ph (B8634^{9 (link)}) with anti-rabbit IgG HRP (Cell Signaling Technology, #7074) and mouse monoclonal anti-VRK1 (Abcam, ab171933), anti-VRK2 (Santa Cruz, sc-365199), anti-H3S10ph (Millipore, 05-806), anti-α-tubulin (B-5-1-2, Sigma, T5168) and anti-MPM2 (Millipore, 05-368) with anti-mouse IgG HRP (Cell Signaling Technology, #7076).

Cartwright T.N., Harris R.J., Meyer S.K., Mon A.M., Watson N.A., Tan C., Marcelot A., Wang F., Zinn-Justin S., Traktman P, & Higgins J.M. (2022). Dissecting the roles of Haspin and VRK1 in histone H3 phosphorylation during mitosis. Scientific Reports, 12, 11210.

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Immunostaining and Western Blotting Protocols

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The following antibodies were used for immunostaining and western blotting:anti-H3K9ac (07–352), anti-H3ac (06–599), anti-H3S10ph (06–570), fluorescein-conjugated goat anti-rabbit IgG (16–237) and anti-H3K4me2 (07–030) were produced by Millipore (Millipore, Billerica, MA, USA). And anti-H4K5ac (ab51997), anti-H3 (ab1791), anti-γH2AX (ab2893) and AP-conjugated goat anti-rabbit IgG (A4187) were purchased from Abcam (Abcam, Cambridge, MA, USA).

Zhang H., Wang P., Hou H., Wen H., Zhou H., Gao F., Wu J., Qiu Z, & Li L. (2016). Histone Modification Is Involved in Okadaic Acid (OA) Induced DNA Damage Response and G2-M Transition Arrest in Maize. PLoS ONE, 11(5), e0155852.

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Immunofluorescence Staining of mESCs

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mESCs were grown on glass coverslips, fixed with 3% formaldehyde in 1× PBS for 10′ at room temperature. Permeabilization was carried out in 0.5% Triton followed by blocking with 1% bovine serum albumin diluted in 1× PBS (Gemini cat 700-110) for 15 min at room temperature. Primary antibody incubation was performed at room temperature for 45 min (Monoclonal ANTI-FLAG^® M2 antibody produced in mouse Millipore-Sigma F1804 at 1/250 dilution), followed by three 5-min washes in 1× PBS, secondary antibody incubation (AlexaFluor594 Goat anti-Mouse IgG Invitrogen A-11005 at 1/10,000 dilution), three 5-min washes in 1× PBS, counterstaining with DAPI, and mounting in 90% glycerol—0.1× PBS—0.1% p-phenylenediamine, pH 9. Images were acquired on a Zeiss spinning disk with 60× objective. In order to avoid loss of loosely attaching mitotic cells for the H3S10 immunostaining in Sororin-AID cells, cells were detached with TryplE, spun in culture medium, resuspended in PBS, and let to attach for 10 min in 1× PBS 25-μl droplets spotted onto 0.1% poly-l-lysine-coated coverslips. Cells were then processed as described above, except that the primary antibody used was Anti-H3S10Ph, rabbit polyclonal, Millipore 05-636.

Nora E.P., Caccianini L., Fudenberg G., So K., Kameswaran V., Nagle A., Uebersohn A., Hajj B., Saux A.L., Coulon A., Mirny L.A., Pollard K.S., Dahan M, & Bruneau B.G. (2020). Molecular basis of CTCF binding polarity in genome folding. Nature Communications, 11, 5612.

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Lysed Cell Protein Analysis Protocol

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Cells were washed once in PBS and then lysed in 141 mM Tris Base, 106 mM Tris HCl, 2% LDS, 1% Benzonase nuclease (E1014, Millipore), pH 8.5, at room temperature for 15 min and snap frozen in liquid nitrogen. The protein concentration was determined using Rapid Gold BCA Protein Assay Kit (Pierce). Cell lysates (20 µg) were run on NuPAGE 4-12% Bis-Tris protein gels (Invitrogen) using standard procedures, transferred to 0.2 µm polyvinylidene fluoride membranes and blocked in 5% milk in TBST (0.05% Tween 20 in Tris-buffered saline) for 1 h at room temperature before overnight incubation with primary antibodies. Membranes were washed thrice in TBST then incubated with a HRP conjugated secondary antibody for 1 h at room temperature in TBST with 5% milk, and signals were detected by chemiluminescence (Clarity ECL, Biorad). Antibodies used in immunoblotting were as follows: rabbit polyclonal anti-Vinculin (Cell Signaling Technology, #13901), anti-Haspin (Abcam, ab21686, lot 156753), and anti-H3T3ph (B8634; (9)) with anti-rabbit IgG HRP (Cell Signaling Technology, #7074) and mouse monoclonal anti-VRK1 (Abcam, ab171933), anti-VRK2 (Santa Cruz, sc-365199), anti-H3S10ph (Millipore, 05-806), anti--tubulin (B-5-1-2, Sigma, T5168) and anti-MPM2 (Millipore, 05-368) with anti-mouse IgG HRP (Cell Signaling Technology, #7076).

Cartwright T.N., Harris R.J., Meyer S.K., Watson N.A., Tan C.C., Wang F., & Higgins J.M. (2021). Dissecting the roles of Haspin and VRK1 in Histone H3 threonine-3 phosphorylation during mitosis.

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