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H3S10p | Histone H3 (p Ser10) (serum)

351 €
AS16 3636 | clonality: polyclonal | host: rabbit | reactivity:human

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AS16 3636

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product information
Background Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Phosphorylation of H3S10 is associated with mitosis.
Immunogen KLH-conjugated synthetic peptide
Host Rabbit
Clonality Polyclonal
Clone
Purity Serum
Format
Quantity 100 ĩl
Reconstitution
Storage Store at -20°C; make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tubes.
Tested applications chromatin immunoprecipiation (ChIP), Dot Blot (Dot), ELISA (ELISA), immunofluorescence (IF), immunoprecipitation (IP), western blot (WB)
Related products AS16 3635-10 | anti-H3S10p | Histone H3 (p Ser10), rabbit antibodies, 10 µg (affinity purified)
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collection of antibodies to epigenetics
Additional information
This antibody preparation contains 0.05% sodium azide and 0.05%.
application information
Recommended dilution 2 µg/million cells (ChIP), 1: 20 000 (Dot), 1: 2000 (IF), 5 µl (IP), 1: 100 (WB)
Expected | apparent MW

15 kDa

Confirmed reactivity

human

Predicted reactivity chicken, D. melanogaster, mouse, plant, rat, Xenopus sp.
Not reactive in No confirmed exceptions from predicted reactivity known at the moment
Additional information To be added when available
Selected references To be added when available, antibody released in March 2016.

application example

ChIP using anti-H3S10p | Histone H3 (p Ser10) (serum) antibodies

ChIP: assays were performed using human HeLa cells treated with colcemid and anti-H3S10p and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit using sheared chromatin from 10 000 cells. A titration of the antibody consisting of 1, 5, and 10 μ l per ChIP experiment was analysed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 μ g/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos and RPL30. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Immunofluorescence using anti-H3S10p | Histone H3 (p Ser10) (serum)


Immunofluorescence:  Hela asynchronous cells were stained anti-H3S10p antibody and with DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. (A) Cells were immunofluorescently labelled with the H3S10p antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight® 488. (B) The nuclei were stained with DAPI, which specifically labels DNA. Phosphorylation of H3 on serine 10 occurs on condensed chromosomes during mitosis.

Immunoprecipitation using anti-H3S10p | Histone H3 (p Ser10) (serum) antibodies
Immunoprecipitation: HeLa cells were treated with colcemid to block the cell cycle in metaphase and were fixed with formaldehyde. Chromatin from 10 000 cells was sheared and used for immunoprecipitation (IP). IP was performed with 5 μl of anti-H3S10p antibodies. The immunoprecipitated proteins were analysed by Western blot with the antibody diluted 1:500 in TBS-Tween containing 5% skimmed milk. Lane 1 shows the result of the IP; a positive control (sheared chromatin from 10 000 cells) and a negative IP control (no antibody added) are shown in lane 2 and 3, respectively.

western blot using anti-H3S10p | Histone H3 (p Ser10) (serum) ani

Western blot: HeLa cells were treated with TSA (figure A) or with colcemid (figure B), and 15 μ g of histone extracts of these cells were analysed by Western blot using anti-H3S10p antibodies diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. The result of the Western analysis with the antibody is shown in lane 1; lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.

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