The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 61 kDa.
Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Use a concentration of 1 µg/ml.
Zyxin binds alpha-actinin and the CRP proteins and is implicated in the regulation and modulation of actin filaments organization, cell adhesion, cell motility, mitosis, and signal transduction. Zyxin may shuttle between focal adhesion plaques and the nucleus. It has been suggested that zyxin is involved in gene transcription by its association with transcription factors in the nucleus.
ICC/IF image of ab58210 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 (ab58210, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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 - Zyxin antibody (ab58210)This image is a courtesy of Anonymous Abreview
Anti-Zyxin antibody (ab58210) at 1/30000 dilution + Lysate prepared from human breast tissue at 10 µg
Secondary HRP-conjugated goat monoclonal to mouse IgG at 1/1000 dilution
Overlay histogram showing HeLa cells stained with ab58210 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab58210, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.