Phospho-Histone H3 (S28) Recombinant Rabbit Monoclonal Antibody [JE56-06]
cat.: HA721795
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Rat, Mouse |
| Applications: | WB, IF-Cell, IHC-P, FC, ChIP |
| Clonality: | Monoclonal |
| Clone number: | JE56-06 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 15 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues surrounding Ser28 of human Histone H3. |
| Positive control: | HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate, PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, HeLa, rat colon tissue. |
| Subcellular location: | Nucleus, Chromosome |
| Recommended Dilutions:
WB IF-Cell IHC-P FC ChIP |
1:1,000 1:10,000 1:2,000 1:1,000 Use 0.5~2 μg for 25 μg of chromatin. |
| Uniprot #: | SwissProt: P68431 Human | P68433 Mouse | Q6LED0 Rat |
| Alternative names: | H3 histone family 3A H3 histone family 3B H3 histone, family 3B (H3.3B) H3.3 H3.3A H3.3B H33_HUMAN H3F3 H3F3A H3f3b Histone H3.3 Histone H3.3Q Histone H3.A Histone H3.B MGC87782 MGC87783 |
Images
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Fig1:
Western blot analysis of Phospho-Histone H3 (S28) on different lysates with Rabbit anti-Phospho-Histone H3 (S28) antibody (HA721795) 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 (HA721795) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of Phospho-Histone H3 (S28) on different lysates with Rabbit anti-Phospho-Histone H3 (S28) antibody (HA721795) at 1/1,000 dilution. Lane 1: PC-12 cell lysate Lane 2: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: PC-12 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: 59 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 (HA721795) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of HeLa cells labeling Phospho-Histone H3 (S28) with Rabbit anti-Phospho-Histone H3 (S28) antibody (HA721795) 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 (HA721795) at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-Phospho-Histone H3 (S28) antibody (HA721795) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA721795) 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. |
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Fig5:
Flow cytometric analysis of HeLa cells labeling Phospho-Histone H3 (S28). Cells were fixed and permeabilized. Then stained with the primary antibody (HA721795, 1μg/mL) (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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig6: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 100ng/mL nocodazole for 18 hours and either Phospho-Histone H3 (S28) (HA721795) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.