Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H3 (mono methyl K4) with ab176877 at 1/1000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of colon tissue is observed. Counter stained with hematoxylin.

Negative control : PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (mono methyl K4) with ab176877 at 1/1000 dilution, followed by goat anti-rabbit Alexa Fluor^® 488 (IgG) (ab150077) secondary antibody at 1/400 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (Alexa Fluor^® 594 goat anti-mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab176877 at 1/1000 dilution followed by ab150120 (goat anti-mouse Alexa Fluor^® 594 secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (goat anti-rabbit Alexa Fluor^® 488; IgG H&L) at 1/400 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).

• ChIP

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ChIP - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with 0.75% formaldehyde for 10min. The ChIP was performed with 25μg of chromatin, 2μg of ab176877 (blue), and 20μl of Anti-rabbit IgG agarose beads. Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the γ-Actin gene (active). Schematic diagram of the γ-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H3 (mono methyl K4) with ab176877 at 1/1000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of colon tissue is observed. Counterstained with hematoxylin.

Negative control : PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H3 (mono methyl K4) with ab176877 at 1/1000 dilution, followed by prediluted Goat Anti-Rabbit IgG H&L (HRP). Nucleus staining on glandular epithelium of colon tissue is observed. Counter stained with hematoxylin.

Negative control : PBS instead of primary ab, secondary ab is prediluted Goat Anti-Rabbit IgG H&L (HRP).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 105 HeLa cells and 2μg of ab176877 [ERP16597]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 μg of ab176877 [ERP16597]. 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.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

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 ab176877 [ERP16597]. 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 ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 4 µg of ab176877. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. 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 containing PBS, BSA, glycerol, and sodium azide (ab176877).

• PepArr

Supplier Data

Peptide Array - Anti-Histone H3 (mono methyl K4) antibody [ERP16597] - ChIP Grade - BSA and Azide free (AB239402)

ab176877 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 Product Protocol section.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab176877).