Anti-STAT6 antibody [YE361] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of bladder tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Clone YE361 (ab215995) has been successfully conjugated by Abcam. This image was generated using Anti-STAT6 antibody [YE361] (Alexa Fluor® 647). Please refer to ab196928 for protocol details.

ab196928 staining STAT6 in HACAT cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 10% normal goat serum in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab196928 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/200 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunocytochemistry/Immunofluorescence analyis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling STAT6 with unpurified ab32520 at 1/100.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling STAT6 with ab32520 at a concentration of 0.79µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 min with DISCOVERY cell conditioning solution (CC1)  100°C, pH 8.5. ab32520 was incubated at 37°C for 16 min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

This IHC data was generated using the same anti-STAT6 antibody clone, YE361, in a different buffer formulation (cat# ab32520).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skin carcinoma tissue labelling STAT6 with unpurified ab32520 at 1/100.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human solitary fibrous tumor tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Intracellular Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling STAT6 with purified ab32520 at 1/30 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemical staining of paraffin embedded human kidney with purified ab32520 at a working dilution of 1/50. 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 (ab32520).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labelling STAT6 with unpurified ab32520 at a dilution of 1/1000. HRP goat anti-rabbit (ab97051) was used at a dilution of 1/500. The antigen retrieval solution was Tris-EDTA buffer, pH 9.0.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Clone YE361 (ab215995) has been successfully conjugated by Abcam. This image was generated using Anti-STAT6 antibody [YE361] (Alexa Fluor® 488). Please refer to ab196478 for protocol details.

ab196478 staining STAT6 in NIH3T3 cells. The cells were fixed with 100% methanol (5 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab196478 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594, shown in red) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

Clone YE361 (ab215995) has been successfully conjugated by Abcam. This image was generated using Anti-STAT6 antibody [YE361] (PE). Please refer to ab223917 for protocol details.

Overlay histogram showing NIH3T3 cells stained with ab223917 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab223917, 1/1000 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

This antibody gave a positive signal in NIH3T3 cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• IP

Unknown

Immunoprecipitation - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

ab32520 (purified) at 1/20 immunoprecipitating STAT6 in 10 μg NIH/3T3 (mouse embyro fibroblast cell line; Lanes 1 and 2, observed at 100 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

All lanes:

Immunoprecipitation - Anti-STAT6 antibody [YE361] (<a href='/en-us/products/primary-antibodies/stat6-antibody-ye361-ab32520'>ab32520</a>)

Predicted band size: 94 kDa

false

• WB

Lab

Western blot - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-STAT6 antibody [YE361] (<a href='/en-us/products/primary-antibodies/stat6-antibody-ye361-ab32520'>ab32520</a>) at 1/20000 dilution

All lanes:

Raji (human Burkitt's lymphoma cell line) cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 94 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

Western blot : Anti-STAT6 antibody [YE361] ab32520 staining at 1000 µg/mL, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 100 kDa in Wild-type A549 cell lysates with no signal observed at this size in STAT6 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-STAT6 antibody [YE361] (<a href='/en-us/products/primary-antibodies/stat6-antibody-ye361-ab32520'>ab32520</a>) at 1000 µg/mL

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human STAT6 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-stat6-knockout-a549-cell-line-ab286716'>ab286716</a>) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 97 kDa

Observed band size: 100 kDa

false

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5µg of ab32520 [YE361]. 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

• WB

Unknown

Western blot - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

All lanes:

Western blot - Anti-STAT6 antibody [YE361] (<a href='/en-us/products/primary-antibodies/stat6-antibody-ye361-ab32520'>ab32520</a>) at 1/1000 dilution

All lanes:

NIH/3T3 (mouse embyro fibroblast cell line) cell lysate

Predicted band size: 94 kDa

Observed band size: 100 kDa

false

• WB

Lab

Western blot - Anti-STAT6 antibody [YE361] - BSA and Azide free (AB215995)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32520).

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-STAT6 antibody [YE361] (<a href='/en-us/products/primary-antibodies/stat6-antibody-ye361-ab32520'>ab32520</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embyro fibroblast cell line) cell lysate at 10 µg

Lane 2:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

Predicted band size: 94 kDa

Observed band size: 100 kDa

false