Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade

• WB

Supplier Data

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 1 µg/mL

Lane 1:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Lane 2:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg

Lane 3:

Histone H4 Recombinant Protein at 0.1 µg

Lane 4:

Histone H3.1 Recombinant Protein at 0.1 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

IHC image of Histone H4 staining in human breast fibroadenoma formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab31380, 0.1μg/ml, for 15 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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

Overlay histogram showing HeLa cells stained with ab31830 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab31830, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

ab31830 staining Histone H4 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab31830 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ChIP

Unknown

ChIP - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

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, 5µg of ab31830 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

• WB

Lab

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 1 µg/mL

Lane 1:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg/mL

Lane 2:

HeLa Nuclear Lysate (Triton enriched) at 10 µg/mL

Lane 3:

Histone H4 Recombinant Protein at 0.1 µg/mL

Lane 4:

Histone H3.1 Recombinant Protein at 0.1 µg/mL

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (AB31830)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade (ab31830) at 5 µg/mL

Lane 1:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg

Lane 2:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg

Lane 3:

Histone H4 Recombinant Protein at 0.1 µg

Lane 4:

Histone H3.1 Recombinant Protein at 0.1 µg

Lane 5:

Histone H2A Recombinant Protein at 0.1 µg

Lane 6:

Histone H2B Recombinant Protein at 0.1 µg

Lane 7:

Histone H1 Recombinant Protein at 0.1 µg

Lane 8:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 9:

HeLa Histone Preparation Nuclear Lysate at 2.5 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 10:

Histone H4 Recombinant Protein at 0.1 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 11:

Histone H3.1 Recombinant Protein at 0.1 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 12:

Histone H2A Recombinant Protein at 0.1 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 13:

Histone H2B Recombinant Protein at 0.1 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Lane 14:

Histone H1 Recombinant Protein at 0.1 µg with Human Histone H4 peptide (<a href='/en-us/products/proteins-peptides/human-histone-h4-peptide-ab13843'>ab13843</a>)

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

false

Exposure time: 2min