Name: Anti-Histone H4 (acetyl K16) antibody [EPR1004]
SKU: ab109463
Rating: 5 (14 reviews)

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

Images

ChIC/CUT&RUN sequencing - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (AB109463), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 2 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

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 -

Tested
Tested

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 -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/200	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

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.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

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.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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15 products for Alternative Product

Target data

See full target information H4C1 acetyl K16

Function

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.

Alternative names

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR1004
Purification technique: Affinity purification Protein A
Specificity: This antibody only detects Histone H4 acetylated on Lysine 16.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

What is this antibody validated in?

Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of Histone H4?

Anti-Histone H4 (acetyl K16) [EPR1004] (ab109463) specifically detects a band for Histone H4 (UniProt: P62805) at a molecular weight of 11kDa.

Recommended positive controls

WB: HeLa, C6 and mouse spleen cell lysates - treated with TSA.IHC-P: Human testis, transitional cell carcinoma and colon tissues.ICC/IF: HeLa cells treated with TSA.Flow Cyt (intra): HeLa cells.ChIC/CUT&RUN-Seq: HeLa cells.

Trusted by the scientific community

Anti-Histone H4 (acetyl K16) [EPR1004] (ab109463) was first used in a scientific publication in 2011 and has been cited over 80 times in peer-reviewed journals.

Reviewed by scientists

Anti-Histone H4 (acetyl K16) [EPR1004] (ab109463) has over 10 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products

We have a range of other formats of antibody clone [EPR1004] also available for your convenience:
ab109463, Carrier free - ab194352, HRP - ab200859, Alexa Fluor® 488 - ab246771, Alexa Fluor® 647 - ab246772

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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.

22 product images

ChIC/CUT&RUN sequencing - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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.
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
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 - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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.
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: HRP-labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L)
Predicted band size: 11 kDa
Observed band size: 11 kDa
Western blot - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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
Secondary
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) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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 - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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 - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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 - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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 - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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).
Flow Cytometry (Intracellular) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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.
Flow Cytometry (Intracellular) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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).
Flow Cytometry (Intracellular) - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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 sequencing - Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)
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.