WB result of Histone H3 (mono methyl K4) Rabbit pAb
Primary antibody: Histone H3 (mono methyl K4) Rabbit pAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: HepG2 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Exposure time: 180 s
Histone H3 (mono methyl K4) Rabbit polyclonal antibody
Histone H3 (mono methyl K4) Rabbit polyclonal antibody
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | H3K4me1 |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P68431 |
| Antibody Type | Polyclonal antibody |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ChIP |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| Dot Blot | 1:1000 | |
| WB | 1:1000 | |
| IHC-P | 1:2000 | |
| ICC | 1:500 | |
| ChIP | 1:20-1:50 |
Background
H3K4me1 is an epigenetic modification to the DNA packaging protein Histone H3. It is a mark that indicates the mono-methylation at the 4th lysine residue of the histone H3 protein and often associated with gene enhancers. H3K4me1 is enriched at active and primed enhancers. Transcriptional enhancers control the cell-identity gene expression and are important in the cell identity.
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Western Blot
WB result of Histone H3 (mono methyl K4) Rabbit pAb
Primary antibody: Histone H3 (mono methyl K4) Rabbit pAb at 1/1000 dilution
Lane 1: Neuro-2a whole cell lysate 20 µg
Lane 2: EL4.IL-2 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Exposure time: 180 s
WB result of Histone H3 (mono methyl K4) Rabbit pAb
Primary antibody: Histone H3 (mono methyl K4) Rabbit pAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Lane 2: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Exposure time: 180 s
Dot Blot
Dot blot result of Histone H3 (mono methyl K4) Rabbit pAb
Lane 1: H3 (mono methyl K4) peptide
Lane 2: H3 (di methyl K4) peptide
Lane 3: H3 (tri methyl K4) peptide
Lane 4: H3 (mono methyl K9) peptide
Lane 5: H3 unmodified peptide
Primary antibody: Histone H3 (mono methyl K4) Rabbit pAb at 1/1000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Exposure time: 18 s
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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-Histone H3 (mono methyl K4) antibody was used at 1/2000 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 stomach. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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 gastric cancer. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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 hepatocellular carcinoma. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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 stomach. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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 colon. Anti-Histone H3 (mono methyl K4) antibody was used at 1/2000 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.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Histone H3 (mono methyl K4) 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).
ChIP
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 (mono methyl K4) Rabbit polyclonal antibody 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 γ-Actin, GAPDH, AFM, MYOD1, SAT-2 and SAT-α in Histone H3 (mono methyl K4) Rabbit polyclonal antibody - immunoprecipitated sample.
