Anti-CD58 antibody [EPR24012-147] - BSA and Azide free (Detector)

• WB

Lab

Western blot - Anti-CD58 antibody [EPR24012-147] - BSA and Azide free (Detector) (AB281051)

This data was developed using ab275392, the same antibody clone in a different buffer formulation.

False colour image of Western blot : Anti-CD58 antibody [EPR24012-147] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab275392 was shown to bind specifically to CD58. A band was observed at 55 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD58 knockout cell line ab265947 (knockout cell lysate ab257880). To generate this image, wild-type and CD58 knockout HeLa 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 (IRDye^� 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^� 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD58 antibody [EPR24012-147] (<a href='/en-us/products/primary-antibodies/cd58-antibody-epr24012-147-ab275392'>ab275392</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 lysate (<a href='/en-us/products/cell-lysates/human-cd58-knockout-hela-cell-lysate-ab257880'>ab257880</a>)

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:

Raji cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 28 kDa

Observed band size: 55 kDa

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• sELISA

Lab

Sandwich ELISA - Anti-CD58 antibody [EPR24012-147] - BSA and Azide free (Detector) (AB281051)

Sandwich ELISA of ab253641 with the capture antibody ab281201 dilution at 2 µg/mL and detector antibody ab281051 dilution at 0.5 µg/mL.