Maltose binding protein fusion protein, corresponding to C terminal amino acids 1866-1996 of Human Separase.
Separase antibodies can be used as a specific marker for centrosomes of mitotic cells. The staining of separase in centrosomes can be detected from prophase of mitosis up until anaphase.
Our Abpromise guarantee covers the use of ab16170 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|WB||Use a concentration of 10 µg/ml. Detects a band of approximately 170 kDa (predicted molecular weight: 233 kDa).|
|ICC/IF||Use at an assay dependent concentration. Used at a dilution of 1/100 for 1 hr on HeLa cells (see Abreview).|
ab16170 stained MCF7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16170 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.