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Synthetic peptide corresponding to C terminal amino acids of Human alpha Tubulin, conjugated to KLH
Our Abpromise guarantee covers the use of ab28439 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|WB||1/2000 - 1/10000. Predicted molecular weight: 54 kDa.|
|Flow Cyt||Use 1µg for 106 cells.|
|ICC/IF||Use a concentration of 10 µg/ml.|
ab28439 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab28439 at 10µg/ml overnight at +4°C. The secondary antibody (green) was anti-mouse DyLight® 488 (ab96879) IgG (H+L) used at a 1/250 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. This antibody also gave a positive result in Methanol fixed (100%) Hek293 and MCF-7 cells, and also in formaldehyde fixed (4%) HeLa, Hek293, HepG2, and MCF-7 cells at 10ug/ml.
Overlay histogram showing HeLa cells stained with ab28439 (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 (ab28439, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was anti-mouse DyLight® 488 (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/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. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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