Anti-STAT2 antibody [22/Stat2] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-STAT2 antibody [22/Stat2] - BSA and Azide free (AB280904)

This data was developed using ab280900 the same antibody clone in a different buffer formulation.

Stat2 was immunoprecipitated from 0.35 mg Ramos (Human Burkitt's lymphoma B lymphocyte), whole cell lysate 10 μg with ab280900 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab280900 at 1/1000 dilution. Mouse IgG for IP (HRP) (ab131368) was used at 1/5000 dilution.

Lane 1 : Ramos whole cell lysate 10 μg

Lane 2 : ab280900 IP in Ramos whole cell lysate

Lane 3 : Mouse monoclonal IgG2a (ab18413) instead of ab280900 in Ramos whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-STAT2 antibody [22/Stat2] (<a href='/en-us/products/primary-antibodies/stat2-antibody-22-stat2-ab280900'>ab280900</a>)

Predicted band size: 97 kDa

Observed band size: 113 kDa,97 kDa

false

• WB

Supplier Data

Western blot - Anti-STAT2 antibody [22/Stat2] - BSA and Azide free (AB280904)

This data was developed using ab280900, the same antibody clone in a different buffer formulation.

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

Exposure time : 92 seconds.

All lanes:

Western blot - Anti-STAT2 antibody [22/Stat2] (<a href='/en-us/products/primary-antibodies/stat2-antibody-22-stat2-ab280900'>ab280900</a>) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 2:

Ramos (Human Burkitt’s lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 3:

THP1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 97 kDa

Observed band size: 113 kDa,97 kDa

false

• WB

Supplier Data

Western blot - Anti-STAT2 antibody [22/Stat2] - BSA and Azide free (AB280904)

This data was developed using ab280900, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1 - 3 : Merged signal (red and green). Green - ab280900 observed at 97, 113 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab280900 Anti-Stat2 antibody was shown to react with STAT2 in HeLa cells in Western blot. Loss of signal was observed when STAT2 knockout sample was used. Wild-type and STAT2 knockout samples were subjected to SDS-PAGE.

ab280900 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-STAT2 antibody [22/Stat2] (<a href='/en-us/products/primary-antibodies/stat2-antibody-22-stat2-ab280900'>ab280900</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

STAT2 knockout HeLa whole cell lysate at 20 µg

Lane 3:

Ramos (Human Burkitt’s lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 97 kDa

Observed band size: 113 kDa,97 kDa

false