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Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Rat ALDH1A1.
(Peptide available as ab25700.)
Our Abpromise guarantee covers the use of ab23375 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||1/200 - 1/400.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-FrFl||Use at an assay dependent concentration.|
|Flow Cyt||Use 0.1µg for 106 cells. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).Can be blocked with Rat ALDH1A1 peptide (ab25700).|
ICC/IF image of ab23375 stained rat PC12 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab23375, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
Overlay histogram showing HepG2 cells stained with ab23375 (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 (ab23375, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (polyclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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