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Synthetic peptide within Human non-muscle Myosin IIA aa 250-350 conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as
Our Abpromise guarantee covers the use of ab75590 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 227 kDa (predicted molecular weight: 227 kDa).|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: non-muscle Myosin IIA knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK-293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab75590 observed at 230 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab75590 was shown to recognize non-muscle Myosin IIA in wild-type HAP1 cells along with additonal cross-reactive bands. No band was observed when non-muscle Myosin IIA knockout samples examined. Wild-type and non-muscle Myosin IIA knockout samples were subjected to SDS-PAGE. ab75590 at a concentration of 1 µg/ml and ab18058 (loading control to Vinculin) at a dilution of 1/1,000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab75590 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab75590 at 5 µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1,000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
ab75590 has not yet been referenced specifically in any publications.
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