WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K27 Rabbit mAb at 1:11000dilution 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 proteinExposure time: 30s.ICC/IF
Confocal imaging of HeLa cells usingTriMethyl-Histone H3-K27 Rabbit mAb(dilution 1:200) followed by a further 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 cells usingTriMethyl-Histone H3-K27 Rabbit mAb(dilution 1:200) followed by a further 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 C6 cells using TriMethylHistone H3-K27 Rabbit mAb (dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG(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 liver tissue usingTriMethyl-Histone H3-K27 Rabbit mAb at a dilution of 1:700 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.IHC
Immunohistochemistry analysis of paraffinembedded Mouse spleen tissue usingTriMethyl-Histone H3-K27 Rabbit mAb at a dilution of 1:700 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.IHC
Immunohistochemistry analysis of paraffinembedded Rat lung tissue using TriMethylHistone H3-K27 Rabbit mAb at adilution of 1:700 (40x lens). High pressureantigen retrieval performed with 0.01M TrisEDTA Buffer (pH 9.0) prior to IHC staining.IP
Immunoprecipitation of TriMethyl-Histone H3-K27 from 600 µg extracts of 293F cells was performed using5 µg of TriMethyl-Histone H3-K27 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-K27 Rabbit mAb at a dilution of 1:20000.CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit(pAG-Tn5) forIllumina from 10⁵ Hela cells with 1μg Tri-MethylHistone H3-K27 Rabbit mAb, alongwith a Goat Anti-Rabbit IgG(H+L). TheCUT&Tag results indicate the enrichmentpattern of H3K27me3 in representative geneloci.CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit(pAG-Tn5) forIllumina from 10⁵ Hela cells with 1μg Tri-MethylHistone H3-K27 Rabbit mAb, alongwith a Goat Anti-Rabbit IgG(H+L). TheCUT&Tag results indicate the enrichmentpattern of H3K27me3 in representative geneloci.ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K27antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitativePCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.| Product Name | TriMethyl-Histone H3-K27 Rabbit mAb |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide. This information is considered to be commercially sensitive. |
| Modification | TriMethyl-K27 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPICC/IFIHCIPWBCUT&Tag-seq |
| WB:1:10000-1:40000 ICC/IF:1:200-1:1000 IHC:1:500- 1:3000 ChIP:5μg antibody for 5μg-10μg of Chromatin IP:0.5μg-4μg antibody for 200μg-600μg extracts of whole cells CUT&Tag:10⁵ cells/1μg |
|
| Species Reactivity | HumanMouseRat |
| 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 | H3t H3.4 H3/g H3FT H3C16 HIST3H3 TriMethyl-Histone H3-K27 |
| Molecular Weight(MW) | 15kDa(Observed band size:17kDa) |
| Cellular Localization | Chromosome, Nucleus. |
WB
Western blot analysis of various lysates using TriMethyl-Histone H3-K27 Rabbit mAb at 1:11000dilution 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 proteinExposure time: 30s.
ICC/IF
Confocal imaging of HeLa cells usingTriMethyl-Histone H3-K27 Rabbit mAb(dilution 1:200) followed by a further 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 cells usingTriMethyl-Histone H3-K27 Rabbit mAb(dilution 1:200) followed by a further 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 C6 cells using TriMethylHistone H3-K27 Rabbit mAb (dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG(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 liver tissue usingTriMethyl-Histone H3-K27 Rabbit mAb at a dilution of 1:700 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.
IHC
Immunohistochemistry analysis of paraffinembedded Mouse spleen tissue usingTriMethyl-Histone H3-K27 Rabbit mAb at a dilution of 1:700 (40x lens).High pressure antigen retrieval performedwith 0.01M Tris-EDTA Buffer (pH 9.0) prior toIHC staining.
IHC
Immunohistochemistry analysis of paraffinembedded Rat lung tissue using TriMethylHistone H3-K27 Rabbit mAb at adilution of 1:700 (40x lens). High pressureantigen retrieval performed with 0.01M TrisEDTA Buffer (pH 9.0) prior to IHC staining.
IP
Immunoprecipitation of TriMethyl-Histone H3-K27 from 600 µg extracts of 293F cells was performed using5 µg of TriMethyl-Histone H3-K27 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-K27 Rabbit mAb at a dilution of 1:20000.
CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit(pAG-Tn5) forIllumina from 10⁵ Hela cells with 1μg Tri-MethylHistone H3-K27 Rabbit mAb, alongwith a Goat Anti-Rabbit IgG(H+L). TheCUT&Tag results indicate the enrichmentpattern of H3K27me3 in representative geneloci.
CUT&Tag
CUT&Tag was performed using the CUT&TagAssay Kit(pAG-Tn5) forIllumina from 10⁵ Hela cells with 1μg Tri-MethylHistone H3-K27 Rabbit mAb, alongwith a Goat Anti-Rabbit IgG(H+L). TheCUT&Tag results indicate the enrichmentpattern of H3K27me3 in representative geneloci.
ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using TriMethyl-Histone H3-K27antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitativePCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.| Application Notes | WB:1:10000-1:40000 ICC/IF:1:200-1:1000 IHC:1:500- 1:3000 ChIP:5μg antibody for 5μg-10μg of Chromatin IP:0.5μg-4μg antibody for 200μg-600μg extracts of whole cells CUT&Tag:10⁵ cells/1μg |
|---|
| 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. |
