Anti-RAB7 antibody [Rab7-117] (ab50533)

  Immunocytochemistry/ Immunofluorescence - RAB7 antibody [Rab7-117] (ab50533)

ICC/IF image of ab50533 stained HepG2 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 (ab50533, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  Western blot - RAB7 antibody [Rab7-117] (ab50533)

Anti-RAB7 antibody [Rab7-117] (ab50533) at 1/1000 dilution + Lysate prepared from mouse neuroblastoma cells at 10 µg

Secondary
HRP-conjugated goat monoclonal to mouse IgG at 1/2000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 23 kDa
Observed band size : 23 kDa


Exposure time : 20 seconds

This image is a courtesy of Anonymous Abreview

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  Western blot - RAB7 antibody [Rab7-117] (ab50533)

Anti-RAB7 antibody [Rab7-117] (ab50533) at 1/500 dilution (for 3 hours at 20°C) + Human brain whole tissue lysate at 15 µg

Secondary
An HRP-conjugated Goat anti-mouse IgG polyclonal at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 23 kDa
Observed band size : 24 kDa (why is the actual band size different from the predicted?)


Exposure time : 1 minute

Blocking Step: 5% Milk for 1 hour at 20°C

This image is courtesy of an anonymous Abreview

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