WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant 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 (tri methyl K4) Recombinant Rabbit mAb (S-4099)
Histone H3 (tri methyl K4) Recombinant Rabbit mAb (S-4099)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Histone H3 (tri methyl K4) |
| Synonyms | H3K4me3 |
| Location | Nucleus |
| Accession | P68431 |
| Clone Number | S-4099 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC, ChIP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | HeLa, NIH/3T3, C6 |
| 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, Rt |
| ICC | 1:500 | Hu, Ms |
| ChIP | 1:20-1:50 | Hu |
Background
Histone H3 lysine 4 trimethylation (commonly denoted as H3K4me3) is a crucial epigenetic modification on the tail of core histone H3 that plays a central role in regulating gene transcription. It is primarily catalyzed by histone methyltransferases from the COMPASS/COMPASS-like family (such as MLL1-4 and SET1A/B) and is specifically recognized by "reader" proteins containing PHD domains (e.g., Taf3 and ING family proteins). The genomic distribution of H3K4me3 is highly characteristic: it is strongly enriched at the promoter regions of actively transcribed genes, particularly near transcription start sites, marking a "primed for activation" state. This modification facilitates the formation of open chromatin structures and the assembly of the transcription initiation complex by recruiting chromatin remodelers, general transcription factors, and histone acetyltransferases, thereby positively regulating gene expression. H3K4me3 is essential for maintaining pluripotency in embryonic stem cells, cell fate determination, and processes such as learning and memory. Notably, its dysregulation—whether excessive or deficient—is closely associated with various developmental disorders and cancers (such as leukemia and breast cancer). Consequently, H3K4me3 is not only a vital biomarker in fundamental research but also a potential target for disease diagnosis and therapy.
Picture
Picture
Western Blot
WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant 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
WB result of Histone H3 (tri methyl K4) Recombinant Rabbit mAb
Primary antibody: Histone H3 (tri methyl K4) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C6 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
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Histone H3 (tri 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).
ICC shows positive staining in NIH/3T3 cells. Anti- Histone H3 (tri 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 (tri methyl K4) Recombinant Rabbit mAb (S-4099) 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 (tri methyl K4) Recombinant
Rabbit mAb (S-4099)-immunoprecipitated sample.
