Anti-Rag D antibody - N-terminal

• IP

Supplier Data

Immunoprecipitation - Anti-Rag D antibody - N-terminal (AB187679)

Immunoprecipitation. ab187679 at 1 μg/ml staining Rag D in 293T whole cell lysate immunoprecipitated using ab187679 at 6 μg/mg lysate (1 mg/IP; 20% of IP loaded/lane).
Lane 2 : control IgG.

All lanes:

Immunoprecipitation - Anti-Rag D antibody - N-terminal (ab187679)

Predicted band size: 46 kDa

true

Exposure time: 3s

• WB

Lab

Western blot - Anti-Rag D antibody - N-terminal (AB187679)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : RRAGD knockout HEK-293T cell lysate 20 ug
Lane 3 : Jurkat cell lysate 20 ug
Lane 4 : HeLa cell lysate 20 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab187679 observed at 48 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab187679 was shown to react with Rag D in wild-type HEK-293T cells in Western blot with loss of signal observed in Rragd knockout cell line ab266287 (Rragd knockout cell lysate ab259091). Wild-type HEK-293T and Rragd knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab187679 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ^°C at a 1 in 2000 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-Rag D antibody - N-terminal (ab187679) at 1/2000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human RRAGD (Rag D) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-rragd-rag-d-knockout-hek-293t-cell-lysate-ab259091'>ab259091</a>) at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 46 kDa

Observed band size: 48 kDa,55 kDa

false

• WB

Supplier Data

Western blot - Anti-Rag D antibody - N-terminal (AB187679)

All lanes:

Western blot - Anti-Rag D antibody - N-terminal (ab187679) at 0.1 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

293T whole cell lysate at 50 µg

Lane 3:

Jurkat whole cell lysate at 50 µg

Lane 4:

TCMK-1 whole cell lysate at 50 µg

Lane 5:

NIH3T3 whole cell lysate at 50 µg

Predicted band size: 46 kDa

true

Exposure time: 3min