Anti-STAT2 antibody [Y141]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STAT2 antibody [Y141] (AB32367)

Intracellular Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling STAT2 with purified ab32367 at 1/60 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT2 antibody [Y141] (AB32367)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STAT2 antibody [Y141] (AB32367)

Immunocytochemistry/ Immunofluorescence analysis of THP-1 (Human monocytic leukemia monocyte) cells labeling STAT2 with purified ab32367 at 1/50 dilution (10 µg/mL). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT2 antibody [Y141] (AB32367)

Immunohistochemical analysis of paraffin-embedded human thyroid cancer using ab32367 (unpurified). Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT2 antibody [Y141] (AB32367)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT2 antibody [Y141] (AB32367)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling STAT2 with purified ab32367 at 1/100 dilution (6.22 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32367 (unpurified) observed at 97 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab32367 was shown to recognize STAT2 in wild-type HAP1 cells as signal was lost at the expected MW in STAT2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and STAT2 knockout samples were subjected to SDS-PAGE. ab32367 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367)

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

STAT2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

K562 whole cell lysate at 20 µg

Lane 4:

THP1 whole cell lysate at 20 µg

Predicted band size: 97 kDa

false

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Blocking/Diluting Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367) at 1/1000 dilution

Lane 1:

Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

Mouse brain lysates at 20 µg

Lane 4:

Rat brain lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 97 kDa

Observed band size: 113 kDa,97 kDa

false

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Lanes 1 - 2 : Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab32367 was shown to react with STAT2 in A549 wild-type cells in western blot with loss of signal observed in STAT2 knockout cell line ab267004 (STAT2 knockout cell lysate ab257183). Wild-type and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk before incubation with ab32367 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

STAT2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human STAT2 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-stat2-knockout-a549-cell-line-ab267004'>ab267004</a>)

Predicted band size: 97 kDa

Observed band size: 97 kDa

false

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Lanes 1- 2 : Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32367 was shown to react with STAT2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267005 (knockout cell lysate ab257184) was used. Wild-type A549 and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

STAT2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human STAT2 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-stat2-knockout-a549-cell-line-ab267005'>ab267005</a>)

Predicted band size: 97 kDa

Observed band size: 97 kDa

false

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Lanes 1- 2 : Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32367 was shown to react with STAT2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout sample ab257185 was used. Wild-type A549 and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

STAT2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human STAT2 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-stat2-knockout-a549-cell-line-ab267006'>ab267006</a>)

Predicted band size: 97 kDa

Observed band size: 97 kDa

false

• WB

Lab

Western blot - Anti-STAT2 antibody [Y141] (AB32367)

Lanes 1- 2 : Merged signal (red and green). Green - ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32367 was shown to react with STAT2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261819 (knockout cell lysate ab257182) was used. Wild-type HeLa and STAT2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [Y141] (ab32367) at 1/5000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

STAT2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human STAT2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-stat2-knockout-hela-cell-line-ab261819'>ab261819</a>)

Predicted band size: 97 kDa

Observed band size: 97 kDa

false