Anti-p53R2 antibody (ab8105) at 1/1000 dilution + U2OS whole cell lysate treated with UV irradiation.

Secondary
Goat anti-rabbit at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 41 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?
Additional bands at: 150 kDa (possible cross reactivity)


Exposure time: 1 minute


Blot was incubated for one hour in 5% milk for blocking, and incubated in primary antibody for 16 hours.

All lanes : Anti-p53R2 antibody (ab8105) at 1 µg/ml

Lane 1 : A431 cell lysate
Lane 2 : 3T3 cell lysate with absence of blocking peptide
Lane 3 : 3T3 cell lysate with presence of blocking peptide

Predicted band size: 41 kDa

ab8105 at 1µg/ml staining p53R2 in human lung tissue by IHC

ICC/IF image of ab8105 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab8105, 10µg/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.

Immunofluorescence of p53R2 in Human Lung cells using ab8105 at 20 ug/ml.