Anti-FE65 antibody [EPR3538] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FE65 antibody [EPR3538] - BSA and Azide free (AB247529)

This data was developed using ab91650, the same antibody clone in a different buffer formulation.

ab91650, at 1/50 dilution, staining FE65 in formalin-fixed, paraffin-embedded Human brain tissue, by Immunohistochemistry.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-FE65 antibody [EPR3538] - BSA and Azide free (AB247529)

This data was developed using ab91650, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-FE65 antibody [EPR3538] (<a href='/en-us/products/primary-antibodies/fe65-antibody-epr3538-ab91650'>ab91650</a>) at 1/2000 dilution

All lanes:

SH-SY5Y cell lysates at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit IgG at 1/1000 dilution

Predicted band size: 77 kDa

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• WB

Lab

Western blot - Anti-FE65 antibody [EPR3538] - BSA and Azide free (AB247529)

False colour image of Western blot : Anti-FE65 antibody [EPR3538] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab91650 was shown to bind specifically to FE65. A band was observed at 65/85 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in APBB1 knockout cell line ab267294 (knockout cell lysate ab257833). To generate this image, wild-type and APBB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-FE65 antibody [EPR3538] (<a href='/en-us/products/primary-antibodies/fe65-antibody-epr3538-ab91650'>ab91650</a>) at 1/2000 dilution

Lane 1:

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

Lane 2:

APBB1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human APBB1 (FE65) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-apbb1-fe65-knockout-hek-293t-cell-line-ab267294'>ab267294</a>)

Lane 3:

SH-SY5Y cell lysate at 20 µg

Lane 4:

A431 cell lysate at 20 µg

Predicted band size: 77 kDa

Observed band size: 65 kDa

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