Name: Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade
SKU: ab32129
Rating: 5 (7 reviews)

Rabbit Recombinant Monoclonal H3 acetyl K9 antibody. Suitable for CUT&Tag, PepArr, IHC-P, IP, ChIP, WB, ICC/IF, ChIP-seq, Flow Cyt (Intra), ChIC/CUT&RUN-seq and reacts with Mouse, Human, Rat samples. Cited in 48 publications.

Images

ChIP - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (AB32129), 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

	CUT&Tag	PepArr	IHC-P	IP	ChIP	WB	ICC/IF	ChIP-seq	Flow Cyt (Intra)	ChIC/CUT&RUN-seq
Human	Expected	Tested	Tested	Tested	Tested	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Tested	Expected	Expected	Tested	Expected	Expected	Expected	Expected
Rat	Expected	Expected	Tested	Expected	Expected	Tested	Expected	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/100	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	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 1/50 - 1/200	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/50 - 1/200	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/50 - 1/200	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	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 2 µg chromatin for 25 µg chromatin	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 Mouse	Dilution info 1/500	Notes -
Species Rat	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	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 4 µg chromatin for 30 µg chromatin	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 -	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 5 µg	Notes -

Expected
Expected

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

Associated Products

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Target data

See full target information H3C1 acetyl K9

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

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, H3FC, HIST1H3C, 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, H3K9ac

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: Y28
Purification technique: Affinity purification Protein A
Specificity: This antibody has weak cross-reactivity with H3R8me1 and H3K4me3 in peptide array. Further optimization may be needed to minimize the cross-reactivity in assays similar to peptide array
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: Avoid freeze / thaw cycle

Notes

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:

- 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.

19 product images

ChIP - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 2 μg of ab32129 (red), and 20 μl of Protein A/G sepharose beads. No antibody 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.
Immunoprecipitation - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
ab32129 (purified) at 1/30 dilution (20 μg/ml) immunoprecipitating Histone H3 acetyl K9 in TSA treated HeLa whole cell lysate.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) treated with 500ng/ml TSA for 4h whole cell lysate 10μg
Lane 2 (+): ab32129 & TSA treated HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32129 in TSA treated HeLa whole cell lysate
For western blotting, ab32129 at 1/500 and veriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Predicted band size: 15 kDa
ChIP-sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 μg of Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129). 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.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Histone H3 with purified ab32129 at 1/250 dilution (4 μg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL) was used as the secondary antibody only control.
This experiment and image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet.
CUT&Tag sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
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 Recombinant Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129) was used, along with a Guinea pig anti-rabbit Secondary. DNA was seg using Illumina NovaSeq S Prime to a depth of 24 million reads.
This image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet.
Flow Cytometry (Intracellular) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab32129 at 1/230 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluorr^® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab32129 [Y28]. 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 University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab32129 [Y28]. 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 University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 5 µg of ab32129 [Y28]. 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 University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
This experiment and image is courtesy of Dr Marek Bartosovic, Gonçalo Castelo-Branco Group, Karolinska Institutet
CUT&Tag - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
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 Recombinant Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129) was used, along with a Guinea pig anti-rabbit Secondary. DNA was seg using Illumina NovaSeq S Prime to a depth of 24 million reads.
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.
ChIP-sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
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 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129). 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.
Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842 was for Histone H3 detection.
All lanes: Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129) at 1/10000 dilution
Lane 1: C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg
Lane 2: C6 (Rat glial tumor glial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysate at 15 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: 15 kDa
Observed band size: 15 kDa
Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842 was for Histone H3 detection.
All lanes: Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129) at 1/10000 dilution
Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysate at 15 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: 15 kDa
Observed band size: 15 kDa
Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842 was for Histone H3 detection.
All lanes: Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129) at 1/10000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysate at 15 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: 15 kDa
Observed band size: 15 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Immunohistochemical analysis of Paraffin-embedded sections human colorectal carcinoma tissue labelling Histone H3 (acetyl K9) with ab32129 at 1/2000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on human colorectal carcinoma tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Immunohistochemical analysis of Paraffin-embedded sections human cerebrum tissue labelling Histone H3 (acetyl K9) with ab32129 at 1/2000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on human cerebrum tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Immunohistochemical analysis of Paraffin-embedded sections mouse colon tissue labelling Histone H3 (acetyl K9) with ab32129 at 1/6000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on mouse colon tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
Immunohistochemical analysis of Paraffin-embedded sections rat colon tissue labelling Histone H3 (acetyl K9) with ab32129 at 1/6000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on rat colon tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Peptide Array - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (ab32129)
ab32129 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.
Please note this antibody has weak cross-reactivity with H3R8me1 and H3K4me3 in peptide array. Further optimization may be needed to minimize the cross-reactivity in assays similar to peptide array.