Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H2A.Z with ab188314 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H2A.Z with ab188314 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows :
-ve control 1 : ab188314 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• ChIP-seq

Lab

ChIP-sequencing - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

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 ab223152 [EPR18090]. ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 10 million sequence tags. The image shows binding across a region of chromosome 1 (RLF gene). ChIP-Seq validation performed by Active Motif, Carlsbad, CA

This image was generated using ab223152. The same clone but a different buffer formulation.

• ChIP

Supplier Data

ChIP - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab188314 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Histone H2A.Z with ab188314 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on rat colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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 H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Histone H2A.Z with ab188314 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on mouse colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• WB

Unknown

Western blot - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (ab188314) at 1/10000 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (ab188314) at 1/10000 dilution

All lanes:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 10s

• PepArr

Supplier Data

Peptide Array - Anti-Histone H2A.Z antibody [EPR18090] - ChIP Grade (AB188314)

ab188314 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.