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Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Human non-muscle Myosin IIA.
(Peptide available as ab99879.)
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: HEK293 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 1µg/ml and ab18058 (loading control to Vinculin) at a dilution of 1/1000 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 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 ab75590 at 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.
ab75590 has not yet been referenced specifically in any publications.
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