Anti-IRAK-1 antibody [EPR18630] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-IRAK-1 antibody [EPR18630] - BSA and Azide free (AB250249)

This data was developed using ab180747, the same antibody clone in a different buffer formulation.

IRAK-1 was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with ab180747 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab180747 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HEK-293 whole cell lysate 10μg (Input).
Lane 2 : ab180747 IP in HEK-293 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab180747 in HEK-293 whole cell lysate.

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

Exposure time : 30 seconds.

The 68kDa band observed is likely a splice variant, according to PMID : 16690127.

All lanes:

Immunoprecipitation - Anti-IRAK-1 antibody [EPR18630] (<a href='/en-us/products/primary-antibodies/irak-1-antibody-epr18630-ab180747'>ab180747</a>)

Predicted band size: 76 kDa

false

• WB

Supplier Data

Western blot - Anti-IRAK-1 antibody [EPR18630] - BSA and Azide free (AB250249)

This data was developed using ab180747, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 10 seconds; Lane 2 : 3 minutes; Lane 3 : 30 seconds; Lane 4 : 1 minute.

There are two splice variants for IRAK-1, IRAK-1b and IRAK-1c, respectively. The product recognizes full length IRAK-1 (80kDa).

All lanes:

Western blot - Anti-IRAK-1 antibody [EPR18630] (<a href='/en-us/products/primary-antibodies/irak-1-antibody-epr18630-ab180747'>ab180747</a>) at 1/1000 dilution

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 3:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 76 kDa

Observed band size: 77 kDa

false

• WB

Lab

Western blot - Anti-IRAK-1 antibody [EPR18630] - BSA and Azide free (AB250249)

This data was developed using ab180747, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)

Lane 2 : IRAK-1 knockout HAP1 whole cell lysate (20 μg)

Lane 3 : HeLa whole cell lysate (20 μg)

Lane 4 : Hek293 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab180747 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab180747 was shown to specifically react with IRAK-1 when IRAK-1 knockout samples were used. Wild-type and IRAK-1 knockout samples were subjected to SDS-PAGE. ab180747 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti-Rabbit and 680CW Goat anti-Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRAK-1 antibody [EPR18630] (<a href='/en-us/products/primary-antibodies/irak-1-antibody-epr18630-ab180747'>ab180747</a>)

Predicted band size: 76 kDa

false