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Recombinant fragment corresponding to Chicken Actin aa 50-70 (N terminal). The immuogen is purified chicken gizzard actin. This is predominantly smooth muscle gamma-actin. Is a pan-actin which recognizes all actin isoforms.
Our Abpromise guarantee covers the use of ab14128 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 - 20 µg/ml. Predicted molecular weight: 43 kDa.
On muscle homogenates subject to SDS-PAGE, reacts relatively uniformly with a 43kDa protein present in skeletal, cardiac, gizzard and aorta tissues. Appears to react with all isoforms of actin found in these preparations and shows a strong reaction with the alpha actin found in skeletal, cardiac and arterial muscle.
Methanol fixed HeLa and NIH/3T3 cells
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab14128 overnight at 4°C. Antibody binding was detected using ab175783 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
ab14128 staining Actin in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab14128 at 5μg/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Mouse Alexa Fluor 488 secondary (ab150117) at 2 μg/ml (shown in green) and Goat anti-Rabbit Alexa Fluor 594 secondary (ab150084) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
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