WB
Western blot analysis of Phospho-Histone H3 (S28) on different lysates with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: HeLa treated with 100ng/mL Nocodazole for 18 hours，then the membrane treated with λpp for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 25 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-Histone H3 (S28) with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/10,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.FC
Flow cytometric analysis of HeLa cells labeling Phospho-Histone H3 (S28). Cells were fixed and permeabilized. Then stained with the primary antibody (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 100ng/mL nocodazole for 18 hours with Phospho-Histone H3 (S28) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Product Name | Phospho-Histone H3 (S28) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phosphopeptide corresponding to residues surrounding Ser28 of human Histone H3. |
| Modification | p-S28 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPFCICC/IFIHCIPWB |
| WB:1:1000 ICC/IF:1:10000 IHC:1:2000 FC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H3C1 |
|---|---|
| Gene Synonyms | H3/A H3FA HIST1H3A |
| Gene Full Name | H3 clustered histone 1 |
| Gene Summary | 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 replication-dependent histone that is 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, Aug 2015] |
| Molecular Weight(MW) | 15 kDa |
| Cellular Localization | Nucleus, Chromosome |
| SwissProt ID | P68431 |
|---|
WB
Western blot analysis of Phospho-Histone H3 (S28) on different lysates with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: HeLa treated with 100ng/mL Nocodazole for 18 hours，then the membrane treated with λpp for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 15 kDa Exposure time: 25 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-Histone H3 (S28) with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/10,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Phospho-Histone H3 (S28) antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
FC
Flow cytometric analysis of HeLa cells labeling Phospho-Histone H3 (S28). Cells were fixed and permeabilized. Then stained with the primary antibody (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 100ng/mL nocodazole for 18 hours with Phospho-Histone H3 (S28) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.| Application Notes | WB:1:1000 ICC/IF:1:10000 IHC:1:2000 FC:1:1000 ChIP:Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
