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咨询Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
参考图片
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling H3K27me3 antibody at 1/500 dilution (0.1 μg) / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Dot blot result of H3K27me3 Rabbit mAb Lane 1: H3K27me3 peptide Lane 2: H3K27me2 peptide Lane 1: H3K27un peptide Lane 2: H3K27me1 peptide Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Exposure time: 90 s
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: HeLa whole cell lysate 20 ug Lane 2: HCT 116 whole cell lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 60 s
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: NIH/3T3 whole cell lysate 20 ug Lane 2: mouse brain lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 120 s
WB result of H3K27me3 Rabbit mAb Primary antibody: H3K27me3 Rabbit mAb at 1/1000 dilution Lane 1: C6 whole cell lysate 20 ug Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 60 s
H3K27me3 Rabbit mAb at 1/50 dilution (1 ug) immunoprecipitating H3K27me3 in 0.4 mg HeLa whole cell lysate. Western blot was performed on the immunoprecipitate using H3K27me3 Rabbit mAb at 1/1000 dilution. Secondary antibody (HRP) for IP was used at 1/400 dilution. Lane 1: HeLa whole cell lysate 20 ug (Input) Lane 2: H3K27me3 Rabbit mAb IP in HeLa whole cell lysate Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate Predicted MW: 17 kDa Observed MW: 17 kDa Exposure time: 90 s
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-H3K27me3 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 rat kidney. Anti-H3K27me3 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 human kidney. Anti-H3K27me3 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 human breast cancer. Anti-H3K27me3 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 human lung squamous cell carcinoma. Anti-H3K27me3 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 malignant peripheral nerve sheath tumor (loss of H3K27me3 expression in partial tumor cells). Anti-H3K27me3 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 nerve sheath tumor. Anti-H3K27me3 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-H3K27me3 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 liver. Anti-H3K27me3 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 rat cerebral cortex. Anti-H3K27me3 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 shows positive staining in HCT116 cells. Anti- H3K27me3 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 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).