Anti-C6 antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-C6 antibody (AB74638)

ICC/IF image of ab74638 stained HeLa cells. The cells were 4% PFA 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 (ab74638, 5μg/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. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa cells at 5μg/ml.

• WB

Project5775****

Western blot - Anti-C6 antibody (AB74638)

C6 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-C6 antibody (ab74638) at 1 µg/mL

Lane 1:

Human Plasma Total Protein Lysate at 2 µg

Lane 2:

Human Plasma Total Protein Lysate at 5 µg

Lane 3:

Human Plasma Total Protein Lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 105 kDa

Observed band size: 110 kDa

false

Exposure time: 2min