Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling STAT3 (phospho Y705) with purified ab76315 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76315).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

This ICC/IF data was generated using the same anti-phospho STAT3 Y705 antibody clone, EP2147Y, in a different buffer formulation (cat# ab76315).

Immunocytochemistry/Immunofluorescence analysis of HeLa +/- IFN-α (50ng/mL, 5 minutes) cells labelling STAT3 (phospho Y705) with ab76315 at 1/500 (4.3 μg/mL). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1500) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue using untreated (left) or alkaline phosphatase-treated (right) labeling STAT3 (phospho Y705) with ab76315 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H& L (HRP) (ab97051) at 1/500 dilution.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76315).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

This data was developed using ab76315, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/mL EGF for 10min (Red) / Untreated control (Green) cells labelling STAT3 with ab76315 at 1/500 dilution (0.1μg) (Red) and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

Tissue Microarrays stained for "Anti-STAT3 (phospho Y705) antibody [EP2147Y]" using "ab76315"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab97051 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab76315 at +4°C overnight followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.

• IP

Unknown

Immunoprecipitation - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

ab76315 (purified) at 1/30 immunoprecipitating STAT3 (phospho Y705) in A431 (Human epidermoid carcinoma cell line) cell lysate treated with EGF. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking/Dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76315).

All lanes:

Immunoprecipitation - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (<a href='/en-us/products/primary-antibodies/stat3-phospho-y705-antibody-ep2147y-ab76315'>ab76315</a>)

Predicted band size: 88 kDa

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• Dot

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Dot Blot - Anti-STAT3 (phospho Y705) antibody [EP2147Y] - BSA and Azide free (AB171358)

Dot blot analysis of STAT3 single phospho peptide pY705 (lane 1) and STAT3 non-phospho peptide (lane 2) with ab76315 at 1/1000. Blocking and dilution buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76315).