WB result of Histone H3 (mono methyl K14) Rabbit mAb
Primary antibody: Histone H3 (mono methyl K14) Rabbit mAb at 1/1000 dilution
Lane 1: HeLa 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
Histone H3 (di methyl K14) Recombinant Rabbit mAb (S-R414)
Histone H3 (di methyl K14) Recombinant Rabbit mAb (S-R414)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | H3K14me2 |
| Location | Nucleus |
| Accession | P68431 |
| Clone Number | S-R414 |
| Antibody Type | Recombinant mAb |
| 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 |
| WB | 1:1000 | Hu, Ms |
| IHC-P | 1:500 | Hu, Ms, Rt |
| ICC | 1:100 | Hu, Ms |
| ChIP | 1:20-1:50 | Hu |
Background
Histone H3 (di-methyl K14) refers to a specific modification of the histone H3 protein, where the lysine residue at position 14 (K14) is di-methylated. Histones are proteins that package and organize DNA into nucleosomes in the nucleus of cells. They play a crucial role in regulating gene expression by controlling the accessibility of DNA to transcription factors and other regulatory proteins. The methylation of histone lysine residues, including K14, is a post-translational modification that can influence chromatin structure and function. Di-methylation of K14 on histone H3 is associated with specific biological processes and can be used as a marker for certain epigenetic states.
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Western Blot
WB result of Histone H3 (mono methyl K14) Rabbit mAb
Primary antibody: Histone H3 (mono methyl K14) Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 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
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti- Histone H3 (di methyl K14) 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. Anti- Histone H3 (di methyl K14) 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 stomach. Anti- Histone H3 (di methyl K14) 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 stomach. Anti- Histone H3 (di methyl K14) 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 lung. Anti- Histone H3 (di methyl K14) 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 testis. Anti- Histone H3 (di methyl K14) 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 lung. Anti- Histone H3 (di methyl K14) 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 testis. Anti- Histone H3 (di methyl K14) 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.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Histone H3 (di methyl K14) antibody was used at 1/100 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).
ICC shows positive staining in NIH/3T3 cells. Anti- Histone H3 (di methyl K14) antibody was used at 1/100 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 (di methyl K14) Recombinant Rabbit mAb (S-R414) 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 and
AFM in Histone H3 (di methyl K14)
Recombinant Rabbit mAb (S-R414)-immunoprecipitated
sample.
