Anti-IRS1 antibody

• WB

Supplier Data

Western blot - Anti-IRS1 antibody (AB245314)

All lanes:

Western blot - Anti-IRS1 antibody (ab245314) at 0.04 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Lane 4:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

Lane 5:

NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 50 µg

Predicted band size: 131 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-IRS1 antibody (AB245314)

Western blot : Anti-IRS1 antibody (ab245314) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta.In Western blot, ab245314 was found to be non-specific. A band was observed at 132 kDa in wild-type A549 cell lysates with no change observed in the IRS1 knockout cell line ab301008. To generate this image, wild-type and IRS1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-IRS1 antibody (ab245314) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

IRS1 knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HCT 116 ab288559 cell lysate at 20 µg

Lane 4:

IRS1 knockout HCT 116 ab301007 cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Lane 6:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 132 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS1 antibody (AB245314)

Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for IRS1 using ab245314 at 1/1000 dilution in immunohistochemical analysis. Detection : DAB staining.

• IP

Supplier Data

Immunoprecipitation - Anti-IRS1 antibody (AB245314)

IRS1 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab245314 at 3 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab245314 at 1 μg/ml.

Lane 1 : ab245314 IP in HeLa whole cell lysate.
Lane 2 : Control IgG IP in HeLa whole cell lysate.

Detection : Chemiluminescence with exposure time of 10 seconds.

All lanes:

Immunoprecipitation - Anti-IRS1 antibody (ab245314)

Predicted band size: 131 kDa

false