PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
12 months from date of receipt / reconstitution, -20 °C as supplied
| 应用 | 稀释度 |
|---|---|
| Dot Blot | 1:1000 |
| WB | 1:1000 |
| IP | 1:50 |
| IHC-P | 1:500 |
| ICC | 1:500 |
| ICFCM | 1:50 |
| ChIP | 1:20~1:50 |
H3K27ac is an epigenetic modification to the DNA packaging protein histone H3. It is a mark that indicates acetylation of the lysine residue at N-terminal position 27 of the histone H3 protein. H3K27ac is associated with the higher activation of transcription and therefore defined as an active enhancer mark. H3K27ac is found at both proximal and distal regions of transcription start site (TSS). H3K27ac is also enriched in the regulatory regions of genes implicated in Alzheimer's disease, including those in tau and amyloid neuropathology.
WB result of Histone H3 (acetyl K27) Rabbit mAb
Primary antibody: Histone H3 (acetyl K27) Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with sodium butyrate (30mM, 4hr) whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 17 kDa
Exposure time: 6 s (This blot was developed with high sensitivity substrate)
WB result of Histone H3 (acetyl K27) Rabbit mAb
Primary antibody: Histone H3 (acetyl K27) Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HeLa treated with Trichostatin A (1μM,18hr) whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 17 kDa
Exposure time: 13 s
WB result of Histone H3 (acetyl K27) Rabbit mAb
Primary antibody: Histone H3 (acetyl K27) Rabbit mAb at 1/1000 dilution
Lane 1: C2C12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 17 kDa
Exposure time: 6 s (This blot was developed with high sensitivity substrate)
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells, treated with 1μM Trichostatin A for 18h (Red) or untreated (Green), labeling Histone H3 (acetyl K27) at 1/50 dilution (1 μg) compared with a rabbit monoclonal IgG isotype control (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Histone H3 (acetyl K27) Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating Histone H3 (acetyl K27) in 0.4 mg HeLa treated with sodium butyrate (30mM, 4hr) whole cell lysate.
Western blot was performed on the immunoprecipitate using Histone H3 (acetyl K27) Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: HeLa treated with sodium butyrate (30mM, 4hr) whole cell lysate 20 µg (Input)
Lane 2: Histone H3 (acetyl K27) Rabbit mAb IP in HeLa treated with sodium butyrate (30mM, 4hr) whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa treated with sodium butyrate (30mM, 4hr) whole cell lysate
Predicted MW: 17 kDa
Observed MW: 17 kDa
Dot blot result of Histone H3 (acetyl K27) Rabbit mAb
Lane 1: H3 (acetyl K27) peptide
Lane 2: H3 unmodified peptide
Primary antibody: Histone H3 (acetyl K27) Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Exposure time: 20 s
IHC shows positive staining in paraffin-embedded human colon. Anti-Histone H3 (acetyl K27) antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse cerebral cortex. Anti-Histone H3 (acetyl K27) antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
ICC analysis of HeLa cells treated with Trichostatin A (1μM,18hr) (top panel) and HeLa cells untreated with Trichostatin A (below panel). Anti- Histone H3 (acetyl K27) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3 (acetyl K27) Recombinant Rabbit mAb (S-699-50) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation. Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA) showed the enrichment of RPL30, GAPDH, MYOD1, AFM, SAT-α and SAT-2 in Histone H3 (acetyl K27) Recombinant Rabbit mAb (S-699-50) - immunoprecipitated sample.
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