Anti-Histone H4 antibody - ChIP Grade

• WB

Unknown

Western blot - Anti-Histone H4 antibody - ChIP Grade (AB7311)

All lanes:

Western blot - Anti-Histone H4 antibody - ChIP Grade (ab7311) at 1/500 dilution

Lane 1:

Calf thymus histone extract at 0.5 µg

Lane 2:

Calf thymus histone extract at 0.5 µg with Human Histone H4 peptide (<a href='/en-us/products/unavailable/human-histone-h4-peptide-ab21044'>ab21044</a>)

Lane 3:

Calf thymus histone extract at 0.5 µg with Histone H4 peptide - acetyl K16 (<a href='/en-us/products/unavailable/histone-h4-peptide-acetyl-k16-ab22376'>ab22376</a>)

Lane 4:

Calf thymus histone extract at 0.5 µg with Human Histone H4 (acetyl K20) peptide (<a href='/en-us/products/unavailable/human-histone-h4-acetyl-k20-peptide-ab20632'>ab20632</a>)

Secondary

All lanes:

Alexa-Fluor Goat anti-rabbit IgG at 1/10000 dilution

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody - ChIP Grade (AB7311)

IHC image of Histone H4 staining in human breast carcinoma FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7311, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 antibody - ChIP Grade (AB7311)

SKN-SH cells were fixed in 4% paraformaldehyde for 10 mins, permeabilized in PBS-0.5% Triton X-100 for 5 mins and incubated for 30 minutes with ab7311 (1/200 dilution). The slides were rinsed once in PBS-Triton (0.1%), twice in PBS then incubated with the secondary antibody for 30 mins. The DNA is stained with DAPI (blue). Clear nuclear staining with ab7311 can be seen (green).

This image is courtesy of Darin McDonald

• IHC-P

AbReview79398****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody - ChIP Grade (AB7311)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of formaldehyde-fixed PBS, Tween-permeabilized human colon cancer tissue. Stained with ab7311 at 1/250 dilution. Secondary antibody used was Agilent Detection System, Rabbit/Mouse HRP. Blocking was done with 5% serum for 15 minutes at 21°C. The sample was incubated with the primary antibody and TBST for 1 hour at 21°C. Antigen retrieval method was heat mediated with EDTA.

This image is courtesy of an anonymous Abreview

• ChIP

Lab

ChIP - Anti-Histone H4 antibody - ChIP Grade (AB7311)

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 25μg of chromatin, 2μg of ab7311 (blue), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• WB

AbReview28969****

Western blot - Anti-Histone H4 antibody - ChIP Grade (AB7311)

All lanes:

Western blot - Anti-Histone H4 antibody - ChIP Grade (ab7311) at 1/500 dilution

All lanes:

Human Bone Whole Tissue Lysate at 25 µg

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Exposure time: 1min

This image is courtesy of an Abreview submitted by Rodrigo Alves.

• WB

CiteAb

Western blot - Anti-Histone H4 antibody - ChIP Grade (AB7311)

Western Blotting using Anti-Histone H4 antibody - ChIP Grade, ab7311. Publication image from Brockdorff, N. et al., 2016, Nat Commun, 27892467. Legend direct from paper.

JARID2 N-terminus mediates direct interaction with H2AK119u1-modified nucleosomes(a) Immunoblot of nucleosomal pull downs from nuclear extract. Total nuclear extract (input) and pull down using streptavidin beads, unmodified nucleosomes or nucleosomes modified by H2AK119u1/H2AK119u1(I44A) were probed with the indicated antibodies using extracts from WT cells (left) or JARID2 KO mESCs (right). Full blots are shown in Supplementary Fig. 9. (b) JARID2 1–530 binds to H2AK119u1-modified mononucleosomes (yellow) more tightly than unmodified (blue) or H2AK199u1(I44A) (red) as monitored by biolayer interferometry (n=3).

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