Anti-STAT1 (phospho Y701) antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-STAT1 (phospho Y701) antibody (AB30645)

ICC/IF image of ab30645 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30645, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho Y701) antibody (AB30645)

Paraffin-embedded human breast carcinoma tissue stained for STAT1 (phospho Y701) using ab30645 at 1/590 dilution in immunohistochemical analysis. Tissue was incubated in the absence (left) or presence (right) of immunizing phospho-peptide.

• IHC-P

AbReview31171****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 (phospho Y701) antibody (AB30645)

ab30645 staining STAT1 (phospho Y701) in murine intestine tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 23°C; antigen retrieval was by heat mediation with a citrate buffer. Samples were incubated with primary antibody (1/400 in Tris Buffer Saline) for 16 hours at 4°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/200) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• WB

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Western blot - Anti-STAT1 (phospho Y701) antibody (AB30645)

Suggested dilution : 1 : 500 - 1 : 1000

All lanes:

Western blot - Anti-STAT1 (phospho Y701) antibody (ab30645)

Lane 1:

Untreated MCF7 lysate (5-30ug).

Lane 2:

EGF treated MCF7 lysate (5-30ug).

Predicted band size: 87 kDa

Observed band size: 47 kDa,87 kDa

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• WB

CiteAb

Western blot - Anti-STAT1 (phospho Y701) antibody (AB30645)

STAT1 (phospho Y701) western blot using anti-STAT1 (phospho Y701) antibody ab30645. Publication image and figure legend from Qiu, X., Fu, Q., et al., 2016, PLoS One, PubMed 26859759.

ab30645 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 ab30645 please see the product overview.

Newcastle disease virus infection impaired IFN-α-induced STAT1 phosphorylation.(A) IFN-α-induced phosphorylated STAT1 degradation in NDV-infected A549 and Vero cells. A549 or Vero cells were infected with NDV strains ZJ1 at MOI 3. At indicated time points post infection, A549 and Vero cells were stimulated with 500 U/ml human IFN-α or IFN-γ in 1 ml DMEM at 37°C for 15 min. Uninfected cells were stimulated with IFN as negative controls (IFNα+/infection-). IFN-α-induced phospho-STAT1 was observed (IFNα+/infection+) for reduction in total STAT1 proteins. IFN-γ-induced phospho-STAT1 proteins were not reduced. (B) Phospho-STAT1 in A549 cells transfected with V-expressing plasmids after stimulation with IFN-α. (C) Expression level of STAT1 and phospho-STAT1 decreased in V-expressing Vero cells after IFN-α stimulation. Vero cells were mock transfected with pCI-neo plasmids. Cells on glass coverlips were transfected with pCI-V/ZJ1 plasmids. At 48 h post-transfection, cells were treated with IFN-α for 15 min prior to fixation as in “Material and methods”. V protein was detected by a mixture of anti-serum Pab-V1 and Pab-V2; STAT1 and phosphorylation were determined by anti-STAT1 antibody (ab31369) and anti-phospho-STAT1 antibody (ab30645). Cellular nuclei were stained with DAPI.

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