Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade is a rabbit recombinant monoclonal antibody that is used to detect Histone H3 in CUT&Tag, ChIC/CUT&RUN-seq, ChIP, ChIP-seq, Dot, Flow cytometry (Intra), ICC/IF, IP, PepArr, Western blot. Suitable for Human, Mouse, Rat samples.
- Recombinant format for unrivaled batch-batch consistency
- Cited in over 50 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
CUT&Tag | Flow Cyt (Intra) | ChIC/CUT&RUN-seq | IP | ChIP | Dot | WB | PepArr | ICC/IF | ChIP-seq | |
---|---|---|---|---|---|---|---|---|---|---|
Human | Expected | Expected | Tested | Expected | Tested | Expected | Tested | Expected | Tested | Tested |
Mouse | Tested | Expected | Expected | Tested | Expected | Expected | Tested | Expected | Tested | Expected |
Rat | Expected | Expected | Expected | Expected | Expected | Expected | Tested | Expected | Expected | Expected |
Synthetic peptide | Not recommended | Not recommended | Not recommended | Not recommended | Not recommended | Tested | Not recommended | Tested | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species Mouse | Dilution info 1/500 | Notes - |
Species Rat | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 2 µg | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 2 µg chromatin for 25.00000 µg chromatin | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes We recommend to use 2% BSA as blocking and antibody dilution buffer. |
Species Rat | Dilution info 1/1000 | Notes We recommend to use 2% BSA as blocking and antibody dilution buffer. |
Species Human | Dilution info 1/1000 | Notes We recommend to use 2% BSA as blocking and antibody dilution buffer. |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info 0.1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/500 | Notes - |
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 4 µg for 30.00000 µg chromatin | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide | Dilution info - | Notes - |
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Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
H3FA, HIST1H3A, H3C2, H3FL, HIST1H3B, H3C3, H3FC HIST1H3C, H3C4, H3FB, HIST1H3D, H3C6, H3FD, HIST1H3E, H3C7, H3FI, HIST1H3F, H3C8, H3FH, HIST1H3G, H3C10, H3FK, HIST1H3H, H3C11, H3FF, HIST1H3I, H3C12, H3FJ, HIST1H3J, H3C1, HIST1H3C, H3FC, Histone H3.1, Histone H3/a, Histone H3/b, Histone H3/c, Histone H3/d, Histone H3/f, Histone H3/h, Histone H3/i, Histone H3/j, Histone H3/k, Histone H3/l, H3K4me3
Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade is a rabbit recombinant monoclonal antibody that is used to detect Histone H3 in CUT&Tag, ChIC/CUT&RUN-seq, ChIP, ChIP-seq, Dot, Flow cytometry (Intra), ICC/IF, IP, PepArr, Western blot. Suitable for Human, Mouse, Rat samples.
- Recombinant format for unrivaled batch-batch consistency
- Cited in over 50 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Chromatin was prepared from Hela (human epithelial cel line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 μg of chromatin, 2 μg of ab213224(red), and 20 μl of Anti-rabbit IgG sepharose beads. 2 μg of Rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 4 μg of Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
CUT&Tag-seq was performed using 200,000 Oli-neu (Oligodendrocyte progenitor) cells. Cells were permeabilized with 0.05% Digitonin and 0.01% NP-40 for 3 minutes. A 1:100 dilution of Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade was used, along with a Guinea pig anti-rabbit Secondary. DNA was seg using Illumina NovaSeq S Prime to a depth of 22 million reads.
Positive control Sox10 chr15:79,091,798-79,329,947 (Mouse mm10 genome used).
This image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet.
Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 μg of chromatin and 4 μg of Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade. ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.
Additional screenshots of mapped reads can be downloaded here.
Histone H3 (tri methyl K4) was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) lysate with ab213224 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213224 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution
Lane 1: NIH/3T3 whole cell lysate (Input).
Lane 2: NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab213224 in NIH/3T3 whole cell lysate.
Exposure time: 3 minutes.
Blocking and dilution buffer and concentration: 5% BSA/TBST.
All lanes: Immunoprecipitation - Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade (ab213224)
Predicted band size: 15 kDa
Observed band size: 15 kDa
ab213224 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Support & downloads section.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1%, Triton X-100-permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Histone H3 (tri methyl K4) with ab213224 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Positive nuclear staining on NIH/3T3 cell lines.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (tri methyl K4) with ab213224 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Positive nuclear staining on HeLa cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Blocking/Dilution buffer: 5% BSA/TBST.
All lanes: Western blot - Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade (ab213224) at 1/5000 dilution
Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 2: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Lane 3: C6 (rat glial tumor cell line) whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 1s
Histone H3 (tri methyl K4) Western blot staining using rabbit Anti-Histone H3 (tri methyl K4) antibody
We recommend to use 2% BSA as blocking and antibody dilution buffer.
All lanes: Western blot - Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade (ab213224) at 1/5000 dilution
All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 15 kDa
Exposure time: 40s
Dot blot analysis of Histone H3 (tri methyl K4) labeled with ab213224 at 1/1000 dilution.
Lane 1: Histone H3K4Me3 peptide.
Lane 2: Histone H3 unmodified peptide.
Lane 3: Histone H3K(18+K36)Me3 peptide.
Lane 4: Histone H3K18Me3 peptide.
Lane 5: Histone H3K4Me2 peptide.
Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution was used as secondary antibody.
Exposure time:3 minutes.
Blocking and dilution buffer: 5% NFDM/TBST.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Histone H3 (tri methyl K4) with ab213224 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 2 µg of ab213224 [EPR20551-225]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 4 µg of ab213224. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
CUT&Tag-seq was performed using 200,000 Oli-neu (Oligodendrocyte progenitor) cells. Cells were permeabilized with 0.05% Digitonin and 0.01% NP-40 for 3 minutes. A 1:100 dilution of Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade was used, along with a Guinea pig anti-rabbit Secondary. DNA was seg using Illumina NovaSeq S Prime to a depth of 22 million reads.
Positive control Sox10 chr15:79,091,798-79,329,947 (Mouse mm10 genome used).
This image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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