Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Immunocytochemical/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling STAT3 (phospho S727) with ab32143 (left) and STAT3 with ab32500 (right) both at a dilution of 1/500.

Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) (1/1000) was used as the secondary antibody (green). DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (1/200) was used as a counterstain (red).

The green staining was increased and translocated from the cytoplasm into the nucleus in the EGF (ab9697 100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signal was decreased. For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Immunohistochemical staining of paraffin embedded human astrocytoma with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Purified ab32143 staining STAT3 (phospho S727) in A431 cells by Immunocytochemistry/ Immunofluorescence. 4% PFA-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labelled with ab32143 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear staining on A431 cell line. The red staining is ab7291 anti-Tubulin (mouse mAb), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Immunohistochemical staining of paraffin embedded rat cerebral cortex with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143). ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 cells (starved overnight and treated with 100ng/ml IL-6 for 30min) and 5 µg of ab32143 [E121-31]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

IHC-P analysis of brain astrocytoma using unpurified ab32143 at 1/50 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).

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

• Dot

Unknown

Dot Blot - Anti-STAT3 (phospho S727) antibody [E121-31] - BSA and Azide free (AB219593)

Dot Blot analysis of Lane 1 : STAT3 (pS727) phospho peptide and Lane 2 : STAT3 non-phospho peptide labeling STAT3 (phospho S727) with ab32143 at 1/1000 dilution (0.009 μg/ml). 5% NFDM /TBST was used as the diluting and blocking buffer and concentration. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100,000 dilution. Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32143).