重组Anti-Histone H3 (acetyl K9)抗体[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 (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 (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.

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 (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 (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.

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 (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 (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.

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 (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 (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.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as GAPDH loading control.

ab176842 was for Histone H3 detection.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as GAPDH loading control.

ab176842 was for Histone H3 detection.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as GAPDH loading control.

ab176842 was for Histone H3 detection.

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 ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

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 ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

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 ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

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.

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.

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.

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 ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. 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.

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 (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) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.

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. 

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.