Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade - BSA and Azide free

17 product images

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  ChIP - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129 (red), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).
  Primers and probes are located in the first kb of the transcribed region.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

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  ChIP-sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation (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 (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.

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  ChIP-sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation (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 (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.

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Histone H3 with purified Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129 at 1/250 dilution (4 μg/mL). Cells were fixed in 100% Methano. 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

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  Flow Cytometry (Intracellular) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

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  Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (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

  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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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 – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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.

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  Peptide Array - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade - BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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.

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  ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade - BSA and Azide free (ab203951)

  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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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 data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

• 

  ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade - BSA and Azide free (ab203951)

  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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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 data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

• 

  ChIC/CUT&RUN sequencing - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade - BSA and Azide free (ab203951)

  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 Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade 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 data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade ab32129).

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  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 – BSA and Azide free (ab203951)

  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 (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.

• 

  Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (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

  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.

• 

  Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade – BSA and Azide free (ab203951)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (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

  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.