WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb at 1:6000dilution incubated at room temperature for 1.5 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Exposure time: 30s.WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb at 1:5000dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Negative control (NC): H3 prokaryotic proteinExposure time: 45s.ICC/IF
Confocal imaging of HeLa cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.ICC/IF
Confocal imaging of NIH/3T3 cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.ICC/IF
Confocal imaging of PC-12 cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.IHC
Immunohistochemistry analysis of paraffinembedded Human colon tissue usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.IHC
Immunohistochemistry analysis of paraffinembedded Mouse esophagus tissue usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.IHC
Immunohistochemistry analysis of paraffinembedded Rat colon tissue using TriMethylHistone H3-K4 Rabbit mAb at adilution of 1:5000 (40x lens). High pressureantigen retrieval performed with 0.01M TrisEDTA Buffer (pH 9.0) prior to IHC staining.DB
Dot-blot analysis of all sorts of peptidesusing TriMethyl-Histone H3-K4 antibody at 1:50000 dilution.IP
Immunoprecipitation of TriMethyl-Histone H3-K4 from 600 µg extracts of HeLa cells was performed using1 µg of TriMethyl-Histone H3-K4 Rabbit mAb. Rabbit IgG isotype control was used toprecipitate the Control IgG sample. IP samples were eluted with 1x reducing Laemmli Buffer. The Inputlane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:2000.ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K4 antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR.Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit (pAG-Tn5) for Illumina from 10⁵ K-562 cells with 1 μg of TriMethylHistone H3-K4 Rabbit mAb,followed by incubation with Goat Anti-RabbitIgG(H+L). The CUT&Tag resultsdenote the enrichment pattern of TriMethylHistone H3-K4 around RPL30 gene.CUT&Tag
UT&Tag was performed using the CUT&TagAssay Kit (pAG-Tn5) for Illumina from 10⁵ K-562 cells with 1 μg of TriMethylHistone H3-K4 Rabbit mAb,followed by incubation with Goat Anti-RabbitIgG (H+L). The CUT&Tag resultsdenote the enrichment pattern of TriMethylHistone H3-K4 across chromosome 8 (upperpanel) and the genomic regionencompassing RPL30, a representative geneenriched in TriMethyl-Histone H3-K4 (lowerpanel).| Product Name | TriMethyl-Histone H3-K4 Rabbit mAb |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide. This information is considered to be commercially sensitive. |
| Modification | TriMethyl-K4 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPDBICC/IFIHCIPWBCUT&Tag-seq |
| WB:1:1000-1:10000 ICC/IF:1:50-1:200 IHC:1:2000-1:8000 ChIP:5μg antibody for 5μg-10μg of Chromatin CUT&Tag:10⁵ cells /1 μg DB:1:10000-1:60000 IP:0.5μg-4μg antibody for 400μg-600μg extracts of whole cells |
|
| Species Reactivity | HumanMouseOtherRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | H3-4 |
|---|---|
| Gene Synonyms | H3t H3.4 H3/g H3FT H3C16 HIST3H3 |
| Gene Full Name | H3.4 histone, cluster member |
| Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq, Aug 2015] |
| Alternative Names | H3.4 H3/g H3FT H3t HIST3H3 Histone H3 HIST1H3A TriMethyl-Histone H3-K4 |
| Molecular Weight(MW) | 15kDa(Observed band size:17kDa) |
| Cellular Localization | Chromosome, nucleus. |
WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb at 1:6000dilution incubated at room temperature for 1.5 hours.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Exposure time: 30s.
WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K4 Rabbit mAb at 1:5000dilution incubated overnight at 4℃.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25 μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit.Negative control (NC): H3 prokaryotic proteinExposure time: 45s.
ICC/IF
Confocal imaging of HeLa cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.
ICC/IF
Confocal imaging of NIH/3T3 cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.
ICC/IF
Confocal imaging of PC-12 cellsusing TriMethyl-Histone H3-K4 Rabbit mAb(dilution 1:200) followed by afurther incubation with Cy3 Goat Anti-RabbitIgG (H+L) (dilution 1:500) (Red). Thecells were counterstained with α-TubulinMouse mAb (dilution 1:400) followedby incubation with ABflo® 488-conjugatedGoat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used fornuclear staining (Blue). Objective: 100x.
IHC
Immunohistochemistry analysis of paraffinembedded Human colon tissue usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.
IHC
Immunohistochemistry analysis of paraffinembedded Mouse esophagus tissue usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.
IHC
Immunohistochemistry analysis of paraffinembedded Rat colon tissue using TriMethylHistone H3-K4 Rabbit mAb at adilution of 1:5000 (40x lens). High pressureantigen retrieval performed with 0.01M TrisEDTA Buffer (pH 9.0) prior to IHC staining.
DB
Dot-blot analysis of all sorts of peptidesusing TriMethyl-Histone H3-K4 antibody at 1:50000 dilution.
IP
Immunoprecipitation of TriMethyl-Histone H3-K4 from 600 µg extracts of HeLa cells was performed using1 µg of TriMethyl-Histone H3-K4 Rabbit mAb. Rabbit IgG isotype control was used toprecipitate the Control IgG sample. IP samples were eluted with 1x reducing Laemmli Buffer. The Inputlane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted usingTriMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:2000.
ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K4 antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR.Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.
CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit (pAG-Tn5) for Illumina from 10⁵ K-562 cells with 1 μg of TriMethylHistone H3-K4 Rabbit mAb,followed by incubation with Goat Anti-RabbitIgG(H+L). The CUT&Tag resultsdenote the enrichment pattern of TriMethylHistone H3-K4 around RPL30 gene.
CUT&Tag
UT&Tag was performed using the CUT&TagAssay Kit (pAG-Tn5) for Illumina from 10⁵ K-562 cells with 1 μg of TriMethylHistone H3-K4 Rabbit mAb,followed by incubation with Goat Anti-RabbitIgG (H+L). The CUT&Tag resultsdenote the enrichment pattern of TriMethylHistone H3-K4 across chromosome 8 (upperpanel) and the genomic regionencompassing RPL30, a representative geneenriched in TriMethyl-Histone H3-K4 (lowerpanel).| Application Notes | WB:1:1000-1:10000 ICC/IF:1:50-1:200 IHC:1:2000-1:8000 ChIP:5μg antibody for 5μg-10μg of Chromatin CUT&Tag:10⁵ cells /1 μg DB:1:10000-1:60000 IP:0.5μg-4μg antibody for 400μg-600μg extracts of whole cells |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20℃. Avoid freeze / thaw cycles. |
| Storage Buffer | PBS containing 50% glycerol and 0.05% BSA, preserved with proclin300 or sodium azide (as specified on the Certificate of Analysis), pH 7.3. |
