Anti-CD58 antibody [EP15041] - BSA and Azide free

• WB

Lab

Western blot - Anti-CD58 antibody [EP15041] - BSA and Azide free (AB251219)

This data was developed using the same antibody clone in a different buffer formulation (ab196648).

Lanes 1-4 : Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control ab8245 observed at 37 kDa.

ab196648 Anti-CD58 antibody [EP15041] was shown to specifically react with CD58 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265947 (knockout cell lysate ab257880) was used. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. ab196648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD58 antibody [EP15041] (<a href='/en-us/products/primary-antibodies/cd58-antibody-ep15041-ab196648'>ab196648</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CD58 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CD58 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cd58-knockout-hela-cell-line-ab265947'>ab265947</a>)

Lane 3:

HAP1 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 28 kDa

Observed band size: 43 kDa

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