Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

ab10799 staining human kidney sections by IHC-P using EXPOSE IHC detection kit (ab80436). Formalin fixed paraffin embedded tissue sections were pre-treated using heat mediated antigen retrieval (using a pressure cooker) with sodium citrate buffer (pH6) for 30 mins. The section was incubated with ab10799, 5μg/ml, for 1 hour at room temperature. DAB was used as the chromogen and the section was counterstained with haematoxylin and mounted with DPX.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

IHC image of Histone H3 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 ab10799, 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

• ChIP

Supplier Data

ChIP - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

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 ab10799 (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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

• WB

Project8616****

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

All lanes:

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - Nuclear Marker and ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-antibody-mabcam-10799-nuclear-marker-and-chip-grade-ab10799'>ab10799</a>) at 1 µg/mL

Lane 1:

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

Lane 2:

Calf Thymus Histone preparation nuclear lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 15 kDa

Observed band size: 17 kDa

false

Exposure time: 90s

• IP

Lab

Immunoprecipitation - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

Histone H3 [mAbcam 10799] - ChIP Grade was immunoprecipitated using 0.5mg HeLa whole cell extract, 5μg of Mouse monoclonal to and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).

The antibody was incubated under agitation with Protein G beads for 10min, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70°C; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10799.

Secondary : Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

Band : 15kDa; [mAbcam 10799] Histone H3 - ChIP Grade.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

All lanes:

Immunoprecipitation - Anti-Histone H3 antibody [mAbcam 10799] - Nuclear Marker and ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-antibody-mabcam-10799-nuclear-marker-and-chip-grade-ab10799'>ab10799</a>)

Predicted band size: 15 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

We recommend using 3% milk as the blocking agent in Western Blot.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab10799).

All lanes:

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - Nuclear Marker and ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-antibody-mabcam-10799-nuclear-marker-and-chip-grade-ab10799'>ab10799</a>) 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 1/5000 dilution

Predicted band size: 15 kDa

true

Exposure time: 4min

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade - BSA and Azide free (AB237967)

This data was developed using ab10799, the same antibody clone in a different buffer formulation

All lanes:

Western blot - Anti-Histone H3 antibody [mAbcam 10799] - Nuclear Marker and ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-antibody-mabcam-10799-nuclear-marker-and-chip-grade-ab10799'>ab10799</a>) at 5 µg/mL

Lane 1:

Histone H2A Recombinant Protein at 0.1 µg/mL

Lane 2:

Histone H2B Recombinant Protein at 0.1 µg/mL

Lane 3:

Histone H3.1 Recombinant Protein at 0.1 µg/mL

Lane 4:

Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg/mL

Lane 5:

HeLa nuclear lysate (triton enriched) at 10 µg/mL

Lane 6:

NIH 3T3 nuclear lysate (triton enriched) at 10 µg/mL

Lane 7:

PC12 nuclear lysate (triton enriched) at 10 µg/mL

Secondary

All lanes:

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

Predicted band size: 15 kDa

false

Exposure time: 20s