Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) is a rabbit monoclonal antibody that is used to detect Histone H4 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
-Over 80 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
ChIC/CUT&RUN-seq | IP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
---|---|---|---|---|---|---|
Human | Tested | Not recommended | Tested | Tested | Tested | Tested |
Mouse | Expected | Not recommended | Tested | Expected | Expected | Expected |
Rat | Expected | Not recommended | Tested | Expected | Expected | Expected |
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 Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes - |
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/2000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/2000 | Notes - |
Species Human | Dilution info 1/1000 - 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/200 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/200 | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/200 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | 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.
H4/A, H4FA, HIST1H4A, H4C2, H4/I, H4FI, HIST1H4B, H4C3, H4/G, H4FG, HIST1H4C, H4C4, H4/B, H4FB, HIST1H4D, H4C5, H4/J, H4FJ, HIST1H4E, H4C6, H4/C, H4FC, HIST1H4F, H4C8, H4/H, H4FH, HIST1H4H, H4C9, H4/M, H4FM, HIST1H4I, H4C11, H4/E, H4FE, HIST1H4J, H4C12, H4/D, H4FD, HIST1H4K, H4C13, H4/K, H4FK, HIST1H4L, H4C14, H4/N, H4F2, H4FN, HIST2H4, HIST2H4A, H4C15, H4/O, H4FO, HIST2H4B, H4C16, H4-16, HIST4H4, H4C1, Histone H4, H4K16ac
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) is a rabbit monoclonal antibody that is used to detect Histone H4 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency
-Over 80 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
This antibody only detects Histone H4 acetylated on Lysine 16.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
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.
CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ChIC/CUT&RUN pAG-MNase ab285373 105 HeLa cells and 2μg of ab109463 [EPR1004]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads.
Additional screenshots of mapped reads can be downloaded here.
Blocking/Diluting buffer and concentration 5% NFDM/TBST
Lanes 1 - 2: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/6000 dilution
Lanes 3 - 4: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/24000 dilution
Lanes 1 and 3: Untreated C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg
Lanes 2 and 4: C6 (Rat glial tumor glial cell) treated with Trichostatin A (final concentration is 500ng/ml) for 4 hours whole cell lysate at 20 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: 11 kDa
Exposure time: 3min
Immunocytochemistry/ Immunofluorescence analysis of untreated HeLa cells (top row) and HeLa+ TSA(500ng/ml, 4h) cells (middle row) labeling Histone H4 (acetyl K16) with ab109463 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488); Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 dilution was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilised with 0.1% tritonX-100. DAPI (blue) was used as a nuclear counterstain.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution
Lane 1: HeLa cell lysates, untreated at 10 µg
Lane 2: HeLa cell lysates treated with TSA at 10 µg
All lanes: HRP-labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution
All lanes: HeLa cell lysate - treated with TSA at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution
All lanes: HeLa cell lysate - treated with TSA at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution
All lanes: C6 cell lysate - treated with TSA at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution
All lanes: C6 cell lysate - treated with TSA at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution
All lanes: Mouse spleen tissue lysate at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L)
Predicted band size: 11 kDa
Observed band size: 11 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution
All lanes: Mouse spleen tissue lysate at 10 µg
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with unpurified ab109463 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with purified ab109463 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Histone H4 with unpurified ab109463 at 1/100.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma labelling Histone H4 (acetly K16) with unpurified ab109463 at 1/100.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/100) and secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/100) and secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/150) and secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/150) and secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Overlay histogram showing HeLa cells stained with unpurified ab109463 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109463, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Intracellular Flow Cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16)with unpurified ab109463 (red) at 1/130. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).
Intracellular Flow Cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/200. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).
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 ab109463 [EPR1004]. 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.
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.
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