Anti-STAT3 (phospho Y705) 抗体 [EP2147Y]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) +/- IFN-α (50 ng/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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

NEAT1 influences STAT3 binding to endocytosis-related genes. The siNEAT1 and siCTRL cells were collected for ChIP assays to analyse the relative fold enrichment of the CAV2 (a), TGFB2 (b), or TGFBR1 promoter (c) with anti-STAT3 or RNAP II antibodies. The data points represent the mean values determined from three independent experiments. The data are presented as the mean ± SD. d The U251 cells were fixed and incubated with anti-STAT3 (red), anti-H3K27Ac (green) or anti-H3K27Cro antibodies (green) before the confocal analysis. The intensity plots for the red and green channels were analysed with the ImageJ software. Scale bars 10 μm. e After transfection with the siNEAT1v2 or negative control siRNA, the U251 cells were fixed and incubated with anti-STAT3 (red) or anti-H3K27Ac antibodies (green) before the confocal analysis. The intensity plots for the red and green channels were analysed with the ImageJ software. Scale bars 10 μm. f After 24 h of incubation with 80 μm crotonyl-CoA or the mock control, the U251 cell lysates were harvested and subjected to an immunoprecipitation assay with anti-STAT3 or anti-IgG antibodies. The retrieval of STAT3, H3K27Ac, H3K27Cro+ and Histone H3 by endogenous STAT3 and IgG was measured by western blotting. *p < 0.001

Image from Wang Z et al., Cell Mol Life Sci. 76(15), Fig 8. Doi: 10.1007/s00018-019-03074-9. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.

• IHC-P

AbReview62872****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded rat colon tissue stained for STAT3 using ab76315 at 1/1000 dilution followed by a Horse radish peroxidase conjugated Goat anti-Rabbit secondary antibody at 1/400 dilution.

Antigen Retrieval : Heat mediated - Heat mediated - Buffer/Enzyme Used : pH 9.0 EDTA

This image was courtesy of an anonymous abReview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labeling STAT3 (phospho Y705) with ab76315 at 1/100 (nuclear staining). Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody. Counterstained with hematoxylin.

• IHC-P

AbReview63092****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded mouse colon tissue stained for STAT3 using ab76315 at 1/1000 dilution followed by a Horse radish peroxidase conjugated Goat anti-Rabbit secondary antibody at 1/400 dilution.

Antigen Retrieval : Heat mediated - Buffer/Enzyme Used : pH 9.0 EDTA 70C 2hr.

This image was courtesy on an anonymous Abreview

• IP

Unknown

Immunoprecipitation - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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.

All lanes:

Immunoprecipitation - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315)

Predicted band size: 88 kDa

false

• WB

Lab

Western blot - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (ab76315) at 1/20000 dilution

Lane 1:

Un-treated HeLa (Human epithelial cell line from cervix adenocarcinoma )cell lysate at 10 µg

Lane 2:

IFN-a-treated HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 88 kDa

Observed band size: 88 kDa

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• Dot

Unknown

Dot Blot - Anti-STAT3 (phospho Y705) antibody [EP2147Y] (AB76315)

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.