ChIP
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa using 5 μg of DiMethyl-Histone H3-K4 Rabbit mAb. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DiMethyl-Histone H3-K4 in the representative genomic region surrounding GAPDH gene.ChIP
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa using 5 μg of DiMethyl-Histone H3-K4 Rabbit mAb. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DiMethyl-Histone H3-K4 across chromosome 12 (upper panel) and the genomic region encompassing GAPDH, a representative gene enriched in DiMethyl-Histone H3-K4 (lower panel).CUT&Tag
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina from 10⁵ HeLa with 1 μg of DiMethyl-Histone H3-K4 Rabbit mAb, followed by incubation with Goat Anti-Rabbit IgG(H+L). The CUT&Tag results denote the enrichment pattern of DiMethyl-Histone H3-K4 across chromosome 12 (upper panel) and the genomic region encompassing GAPDH, a representative gene enriched in DiMethyl-Histone H3-K4 (lower panel).CUT&Tag
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina from 10⁵ HeLa cells with 1 μg of DiMethyl-Histone H3-K4 Rabbit mAb, followed by incubation with Goat Anti-Rabbit IgG(H+L. The CUT&Tag results denote the enrichment pattern of DiMethyl-Histone H3-K4 around GAPDH gene.WB
Western blot analysis of various lysates using DiMethyl-Histone H3-K4 Rabbit mAb at 1:3000 dilution 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 protein Exposure time: 30 s.DB
Dot-blot analysis of all sorts of peptides using DiMethyl-Histone H3-K4 Rabbit mAb at 1:5000 dilution.IHC
Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.IHC
Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.IHC
Immunohistochemistry analysis of paraffin-embedded Rat pancreas tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.ICC/IF
Confocal imaging of HeLa cells using DiMethyl-Histone H3-K4 Rabbit pAb (dilution 1:500) followed by a further incubation with Cy3-conjugated Goat anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.IP
Immunoprecipitation of DiMethyl-Histone H3-K4 from 600 µg extracts of HeLa cells was performed using 3 µg of DiMethyl-Histone H3-K4 Rabbit mAb. Rabbit Control IgG was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:10000.ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using DiMethyl-Histone H3-K4 Rabbit mAb 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.| Product Name | DiMethyl-Histone H3-K4 Rabbit mAb |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide. This information is considered to be commercially sensitive. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPDBICC/IFIHCIPWBCUT&Tag-seq |
| WB:1:2000-1:10000 IP:0.5μg-5μg antibody for 400μg-600μg extracts of whole cells ICC/IF:1:200-1:1000 IHC:1:3000-1:12000 DB:1:2000-1:6000 ChIP:5μg antibody for 5μg-10μg of Chromatin ChIP-seq:1:50-1:200 CUT&Tag:10⁵cells/1μg |
|
| 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] |
| Molecular Weight(MW) | 15 kDa(Observed band size: 17 kDa) |
| Cellular Localization | Chromosome, nucleus. |
| SwissProt ID | P68431 |
|---|
ChIP
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa using 5 μg of DiMethyl-Histone H3-K4 Rabbit mAb. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DiMethyl-Histone H3-K4 in the representative genomic region surrounding GAPDH gene.
ChIP
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa using 5 μg of DiMethyl-Histone H3-K4 Rabbit mAb. DNA libraries were prepared using Scale ssDNA-seq Lib Prep Kit for Illumina V2. The ChIP sequencing results indicate the enrichment pattern of DiMethyl-Histone H3-K4 across chromosome 12 (upper panel) and the genomic region encompassing GAPDH, a representative gene enriched in DiMethyl-Histone H3-K4 (lower panel).
CUT&Tag
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina from 10⁵ HeLa with 1 μg of DiMethyl-Histone H3-K4 Rabbit mAb, followed by incubation with Goat Anti-Rabbit IgG(H+L). The CUT&Tag results denote the enrichment pattern of DiMethyl-Histone H3-K4 across chromosome 12 (upper panel) and the genomic region encompassing GAPDH, a representative gene enriched in DiMethyl-Histone H3-K4 (lower panel).
CUT&Tag
CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina from 10⁵ HeLa cells with 1 μg of DiMethyl-Histone H3-K4 Rabbit mAb, followed by incubation with Goat Anti-Rabbit IgG(H+L. The CUT&Tag results denote the enrichment pattern of DiMethyl-Histone H3-K4 around GAPDH gene.
WB
Western blot analysis of various lysates using DiMethyl-Histone H3-K4 Rabbit mAb at 1:3000 dilution 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 protein Exposure time: 30 s.
DB
Dot-blot analysis of all sorts of peptides using DiMethyl-Histone H3-K4 Rabbit mAb at 1:5000 dilution.
IHC
Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
IHC
Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
IHC
Immunohistochemistry analysis of paraffin-embedded Rat pancreas tissue using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:5000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
ICC/IF
Confocal imaging of HeLa cells using DiMethyl-Histone H3-K4 Rabbit pAb (dilution 1:500) followed by a further incubation with Cy3-conjugated Goat anti-Rabbit IgG (H+L) (dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.
IP
Immunoprecipitation of DiMethyl-Histone H3-K4 from 600 µg extracts of HeLa cells was performed using 3 µg of DiMethyl-Histone H3-K4 Rabbit mAb. Rabbit Control IgG was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using DiMethyl-Histone H3-K4 Rabbit mAb at a dilution of 1:10000.
ChIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using DiMethyl-Histone H3-K4 Rabbit mAb 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.| Application Notes | WB:1:2000-1:10000 IP:0.5μg-5μg antibody for 400μg-600μg extracts of whole cells ICC/IF:1:200-1:1000 IHC:1:3000-1:12000 DB:1:2000-1:6000 ChIP:5μg antibody for 5μg-10μg of Chromatin ChIP-seq:1:50-1:200 CUT&Tag:10⁵cells/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. |
