Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

IHC image of ab17036 staining in Human Hodgekin's Lymphoma 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 ab17036, 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.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

IHC image of Histone H4 staining in formalin fixed, paraffin embedded human Hodgkin's lymphoma tissue section*, performed on a Leica Bond™ 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 ab17036, 1 μg/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.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ChIP

Unknown

ChIP - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

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 ab17036 (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

Project

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) 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

Secondary

All lanes:

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

false

Exposure time: 1min

• WB

Project

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 5 µg/mL

Lane 1:

Histone H1 Recombinant Protein at 0.1 µg

Lane 2:

Histone H2A Recombinant Protein at 0.1 µg

Lane 3:

Histone H2B Recombinant Protein at 0.1 µg

Lane 4:

Histone H3.1 Recombinant Protein at 0.1 µg

Lane 5:

Histone H4 Recombinant Protein at 0.1 µg

Secondary

All lanes:

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

false

Exposure time: 30s

• WB

Unknown

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 1 µg/mL

Lane 1:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

true

Exposure time: 1min

• WB

Lab

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

All lanes:

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 2 µg/mL

Lane 1:

Histone H1 Recombinant Protein at 0.1 µg

Lane 2:

Histone H2A Recombinant Protein at 0.1 µg

Lane 3:

Histone H2B Recombinant Protein at 0.1 µg

Lane 4:

Histone H3.1 Recombinant Protein at 0.1 µg

Lane 5:

Histone H4 Recombinant Protein at 0.1 µg

Secondary

All lanes:

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

false

Exposure time: 30s

• WB

CiteAb

Western blot - Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (AB17036)

Histone H4 western blot using anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade ab17036. Publication image and figure legend from Dang, G., Cui, Y., et al., 2018, Front Immunol, PubMed 29670633.

ab17036 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab17036 please see the product overview.

Recombinant Rv0888NS/MS sphingomyelinase induces neutrophil extracellular traps (NETs) formation in lung. (A) Immunofluorescence staining of lung sections from mice 48 h after intraperitoneal injection of recombinant Rv0888NS/MS, D438A/MS, and H481N/MS (n = 3 mice per group) administration was performed and compared with the control section PBS and recombinant pMV262/MS (n = 3 mice per group) for DNA (blue), CitH3 (green), and myeloperoxidase (MPO) (red). Images are representative of one of three independent experiments with three mice per group. The recombinant Rv0888NS/MS and D438A/MS groups' merge showed co-localization of MPO with CitH3, indicating NETs' formation. Scale bars = 50 μm. The higher magnification views of the insets (1 and 2), which were randomly chosen, show co-localization of DNA (blue), CitH3 (green), and MPO (red) in NETs structures. Scale bars = 10 μm. (B) Western blot analysis of MPO, histone H4, and CitH3 protein levels in the bronchoalveolar lavage fluid. Images are representative of one of three independent experiments with three mice per group. (C) Reactive oxygen species (ROS) were measured in the serum and lung homogenates of infected mice using a Mouse ROS enzyme-linked immunosorbent assay (ELISA) Kit. (D) Ceramide in the serum and lung homogenates of infected mice was determined by the Mouse Ceramide ELISA Kit. The error bars show the SEM of three independent experiments with three mice per group. (C,D) One-way ANOVA followed by Bonferroni's multiple comparison post hoc test, *p < 0.05, **p < 0.01, ***p < 0.001, ns, non-significant.

false