Anti-Flotillin 1 antibody [EP446Y]

• WB

Lab

Western blot - Anti-Flotillin 1 antibody [EP446Y] (AB78178)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/2000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

A431 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 49 kDa

false

• WB

Unknown

Western blot - Anti-Flotillin 1 antibody [EP446Y] (AB78178)

Lanes 1-4 : Merged signal (red and green). Green - ab78178 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab78178 Anti-Flotillin 1 antibody [EP446Y] was shown to specifically react with Flotillin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267276 (knockout cell lysate ab257109) was used. Wild-type and Flotillin knockout samples were subjected to SDS-PAGE. ab78178 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-Flotillin 1 antibody [EP446Y] (ab78178) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

FLOT1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-flot1-flotillin-1-knockout-hek-293t-cell-line-ab267276'>ab267276</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HAP1 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: 47 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-Flotillin 1 antibody [EP446Y] (AB78178)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Flotillin 1 knockout HAP1 whole cell lysate (20 μg)

Lanes 1 - 2 : Merged signal (red and green). Green - ab78178 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab78178 was shown to specifically react with Flotillin 1 in wild-type HAP1 cells as signal was lost in Flotillin 1 knockout cells. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab78178 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178)

Predicted band size: 47 kDa

false

• WB

Lab

Western blot - Anti-Flotillin 1 antibody [EP446Y] (AB78178)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/2000 dilution

Lane 1:

mouse brain lysate at 10 µg

Lane 2:

rat brain lysate at 20 µg

Lane 3:

C6 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 49 kDa

false

• WB

Unknown

Western blot - Anti-Flotillin 1 antibody [EP446Y] (AB78178)

All lanes:

Western blot - Anti-Flotillin 1 antibody [EP446Y] (ab78178) at 1/500 dilution

All lanes:

K562 cell lysate

Predicted band size: 47 kDa

Observed band size: 48 kDa

false