Application
Dot blot analysis of peptides containing modifications of histone H3 and H4 and unmodified histone H3 using H3S10ph antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane for analysis.Application
WB analysis of histone extracts (15 μg) from TSA-treated HeLa cells using H3S10ph antibody at a dilution of 1:500 in TBS-Tween containing 5% skimmed milk (lane 1). Lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.Application
WB analysis of histone extracts (15 μg) from colcemid-treated HeLa cells using H3S10ph antibody at a dilution of 1:500 in TBS-Tween containing 5% skimmed milk (lane 1). Lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.Application
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 H3S10ph antibody. The immunoprecipitated proteins were analysed by WB with H3S10ph antibody at a dilution of 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.Application
ChIP was performed with colcemid-treated HeLa extracts and either control IgG (5 μg) or Histone H3S10ph (phospho Ser10) antibody (1, 5, and 10 μl per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to the promoter of the active genes c-fos and RPL30. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature.Application
ELISA was performed using a serial dilution of Histone H3S10ph (phospho Ser10) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:35,000.Application
ICC/IF analysis of HeLa asynchronous cells using Histone H3S10ph (phospho Ser10) antibody (green) and DAPI (blue). Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. The cells were immunofluorescently labelled with Histone H3S10ph (phospho Ser10) antibody at a dilution of 1:200.| Product Name | Histone H3S10ph (phospho Ser10) antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to H3 (phospho Ser10)Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq, Jul 2008] |
| Immunogen | Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (H3S10p), using a KLH-conjugated synthetic peptide. |
| Modification | p-Ser10 |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | DotELISAICC/IFIPWBChIP |
| ChIP (1 µl/ChIP), ELISA (1:1,000 – 1:5,000), Dot blotting (1:20,000), Western blotting (1:500), Immunofluorescence (1:200), Immunoprecipitation (5 µl/IP): |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Gene Synonyms | histone cluster 1 H3a H3/A HIST1H3A H3FA |
|---|---|
| Target | H3 (phospho Ser10) |
Application
Dot blot analysis of peptides containing modifications of histone H3 and H4 and unmodified histone H3 using H3S10ph antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane for analysis.
Application
WB analysis of histone extracts (15 μg) from TSA-treated HeLa cells using H3S10ph antibody at a dilution of 1:500 in TBS-Tween containing 5% skimmed milk (lane 1). Lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.
Application
WB analysis of histone extracts (15 μg) from colcemid-treated HeLa cells using H3S10ph antibody at a dilution of 1:500 in TBS-Tween containing 5% skimmed milk (lane 1). Lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.
Application
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 H3S10ph antibody. The immunoprecipitated proteins were analysed by WB with H3S10ph antibody at a dilution of 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.
Application
ChIP was performed with colcemid-treated HeLa extracts and either control IgG (5 μg) or Histone H3S10ph (phospho Ser10) antibody (1, 5, and 10 μl per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to the promoter of the active genes c-fos and RPL30. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature.
Application
ELISA was performed using a serial dilution of Histone H3S10ph (phospho Ser10) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:35,000.
Application
ICC/IF analysis of HeLa asynchronous cells using Histone H3S10ph (phospho Ser10) antibody (green) and DAPI (blue). Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. The cells were immunofluorescently labelled with Histone H3S10ph (phospho Ser10) antibody at a dilution of 1:200.| Application Notes | ChIP (1 µl/ChIP), ELISA (1:1,000 – 1:5,000), Dot blotting (1:20,000), Western blotting (1:500), Immunofluorescence (1:200), Immunoprecipitation (5 µl/IP): |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | Whole antiserum from rabbit containing 0.05% azide. |
