Anti-Histone H3 (phospho T3) antibody [EP1702Y] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H3 (phospho T3) antibody [EP1702Y] - BSA and Azide free (AB247494)

This data was developed using ab78351, the same antibody clone in a different buffer formulation.

Overlay histogram showing HeLa cells stained with ab78351 (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 (ab78351, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (IgG; H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T3) antibody [EP1702Y] - BSA and Azide free (AB247494)

This data was developed using the same antibody clone in a different buffer formulation (ab78351).
Immunocytochemistry analysis of HeLa (human cervix adenocarcinoma epithelial cell) labeling Histone H3 with purified ab78351 at 1/250 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.18 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control : PBS instead of the primary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho T3) antibody [EP1702Y] - BSA and Azide free (AB247494)

This data was developed using ab78351, the same antibody clone in a different buffer formulation.

ab78351 at 1/100 dilution staining Histone H3 in human breast carcinoma tissue, using a HRP/AP polymerized secondary antibody.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

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Western blot - Anti-Histone H3 (phospho T3) antibody [EP1702Y] - BSA and Azide free (AB247494)

This data was developed using ab78351, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Histone H3 (phospho T3) antibody [EP1702Y] (<a href='/en-us/products/primary-antibodies/histone-h3-phospho-t3-antibody-ep1702y-ab78351'>ab78351</a>) at 1/500000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HeLa cell lysate treated with FBS + Calyculin A. at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 15 kDa

Observed band size: 14 kDa

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