Anti-STAT1 antibody [1/Stat1] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (AB282026)

This data was developed using ab281999 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling STAT1 with ab281999 at 1/2000 (0.511 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human tonsil. The section was incubated with ab281999 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (AB282026)

This data was developed using ab281999 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling STAT1 with ab281999 at 1/2000 (0.511 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in mouse spleen.The section was incubated with ab281999 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (AB282026)

This data was developed using ab281999 the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized treated and untreated RAW 264.7 cells labelling STAT1 with ab281999 at 1/25 (40.88 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in RAW 264.7 cells treated with IFN-y (10 ng/ml) for 30 min. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• WB

Lab

Western blot - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (AB282026)

This data was developed using ab281999 the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure times : Lanes 1-2 : 37 seconds ;

Lanes 3-4 : 26 seconds.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 19478064, 26859759).

All lanes:

Western blot - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (ab282026) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HeLa treated with 10 ng/ml IFN-gamma for 30 minutes, whole cell lysate at 10 µg

Lane 3:

Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 4:

RAW264.7 treated with 50 ng/ml IFN-gamma for 24 hours, whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 87 kDa

Observed band size: 87 kDa

false

• WB

Lab

Western blot - Anti-STAT1 antibody [1/Stat1] - BSA and Azide free (AB282026)

This data was developed using ab281999, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : Lanes 1-6 : 37 seconds ;

Lanes 7-9 : 3 minutes.

All lanes:

Western blot - Anti-STAT1 antibody [1/Stat1] (<a href='/en-us/products/primary-antibodies/stat1-antibody-1-stat1-ab281999'>ab281999</a>) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Lane 6:

Human colon cancer lysate at 10 µg

Lane 7:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 8:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Lane 9:

C2C12 (mouse myoblasts myoblast), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 87 kDa

Observed band size: 87 kDa

false