Anti-Flotillin 1 antibody

• WB

Project3006****

Western blot - Anti-Flotillin 1 antibody (AB41927)

All lanes:

Western blot - Anti-Flotillin 1 antibody (ab41927) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

Predicted band size: 47 kDa

Observed band size: 35 kDa,50 kDa

true

Exposure time: 4min

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody (AB41927)

ICC/IF image of ab41927 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab41927 at 1μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

• ICC/IF

AbReview9925****

Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody (AB41927)

ab41927 (1/200) staining Flotillin 1 in HeLa cells (green). Cells were fixed with Methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

This image is part of an abreview submitted by Dr. Kirk McManus

• WB

Lab

Western blot - Anti-Flotillin 1 antibody (AB41927)

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

Lanes 1 - 4 : Merged signal (red and green). Green - ab41927 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab41927 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. ab41927 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml 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 (ab41927)

Predicted band size: 47 kDa

false

• WB

Project3006****

Western blot - Anti-Flotillin 1 antibody (AB41927)

All lanes:

Western blot - Anti-Flotillin 1 antibody (ab41927) at 1 µg/mL

Lane 1:

Lung (Human) Tissue Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

false