Anti-CD40 antibody [41/CD40] - BSA and Azide free

• WB

Lab

Western blot - Anti-CD40 antibody [41/CD40] - BSA and Azide free (AB280214)

This data was developed using the same antibody clone in a different buffer formulation (ab280207).

Lanes 1 - 4 : Merged signal (red and green). Green - ab280207 observed at 45 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab280207 was shown to react with CD40 in wild-type U-2 OS cells in Western blot with loss of signal observed in CD40 knockout cell line ab262486 (CD40 knockout cell lysate ab263923). Wild-type U-2 OS and CD40 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab280207 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD40 antibody [41/CD40] (<a href='/en-us/products/primary-antibodies/cd40-antibody-41-cd40-ab280207'>ab280207</a>) at 1/1000 dilution

Lane 1:

Wild-type U-2 OS Vehicle Control LPS (0µg/mL, 6h) cell lysate at 20 µg

Lane 2:

Wild-type U-2 OS Treated LPS (1µg/mL, 6h) cell lysate at 20 µg

Lane 3:

CD40 knockout U-2 OS Vehicle Control LPS (0µg/mL, 6h) cell lysate at 20 µg

Lanes 3 - 4:

Western blot - Human CD40 knockout U-2 OS cell lysate (<a href='/en-us/products/cell-lysates/human-cd40-knockout-u-2-os-cell-lysate-ab263923'>ab263923</a>) at 20 µg

Lane 4:

CD40 knockout U-2 OS Treated LPS (1µg/mL, 6h) cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-CD40 antibody [41/CD40] - BSA and Azide free (AB280214)

This data was developed using ab280207, the same antibody clone in a different buffer formulation.

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

Negative control : Jurkat (PMID : 10498643).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18312851).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-CD40 antibody [41/CD40] (<a href='/en-us/products/primary-antibodies/cd40-antibody-41-cd40-ab280207'>ab280207</a>) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 10 µg

Lane 2:

Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate at 10 µg

Lane 3:

Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 10 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 35 kDa,44 kDa

false

• WB

Supplier Data

Western blot - Anti-CD40 antibody [41/CD40] - BSA and Azide free (AB280214)

This data was developed using ab280207, the same antibody clone in a different buffer formulation.

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18312851).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-CD40 antibody [41/CD40] (<a href='/en-us/products/primary-antibodies/cd40-antibody-41-cd40-ab280207'>ab280207</a>) at 1/1000 dilution

Lane 1:

Human tonsil lysates at 10 µg

Lane 2:

Human lymph node lysates at 10 µg

Lane 3:

Human lymphoma lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 35 kDa,44 kDa

false