Anti-Flotillin 2/ESA antibody [EPR14128(B)]

• IP

Supplier Data

Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (AB181988)

Immunoprecipitation of Hela cell lysate labeling Flotillin 2/ESA using ab181988 at a 1/40 dilution (lane 1). Lane 2 shows the negative control. Anti-Rabbit IgG (HRP) specific to the non-reduced form of IgG, was used as a secondary antibody at a dilution of 1/1500. Blocking/dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)

Predicted band size: 47 kDa

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• WB

Lab

Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (AB181988)

Lanes 1-4 : Merged signal (red and green). Green - ab181988 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa.

ab181988 Anti-Flotillin 2/ESA antibody [EPR14128(B)] was shown to specifically react with Flotillin 2/ESA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266847 (knockout cell lysate ab257442) was used. Wild-type and Flotillin 2/ESA knockout samples were subjected to SDS-PAGE. ab181988 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

FLOT2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-flot2-flotillin-2-esa-knockout-hek-293t-cell-line-ab266847'>ab266847</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A549 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 1/10000 dilution

Predicted band size: 27 kDa,47 kDa

Observed band size: 27 kDa,47 kDa

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• WB

Supplier Data

Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (AB181988)

Blocking buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988) at 1/4000 dilution

Lane 1:

293 cell lysate at 20 µg

Lane 2:

Hela cell lysate at 20 µg

Lane 3:

A375 cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 47 kDa

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