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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human CLK2.
(Peptide available as ab99082.)
Our Abpromise guarantee covers the use of ab86147 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.5 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).|
|ICC/IF||Use a concentration of 5 µg/ml.|
ab86147 staining CLK2 in human colon carcinoma cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol and blocked with 1% BSA for 30 minutes at 37°C. Samples were incubated with primary antibody (1/30 in PBS + 1% BSA) for 16 hour at 4°C. A FITC-conjugated goat anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.
ICC/IF image of ab86147 stained HepG2 cells. The cells were 4% PFA 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 ab86147 at 5ug 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. This antibody also gave a positive result in HeLa pfa fixed cell types at 5ug/ml