The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.5 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).
Use a concentration of 5 µg/ml.
Phosphorylates serine- and arginine-rich (SR) proteins of the spliceosomal complex may be a constituent of a network of regulatory mechanisms that enable SR proteins to control RNA splicing. Phosphorylates serines, threonines and tyrosines.
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. Lammer subfamily. Contains 1 protein kinase domain.
Autophosphorylates on all three types of residues.
Immunocytochemistry/ Immunofluorescence - Anti-CLK2 antibody (ab86147)This image is courtesy of an Abreview submitted by Eleni Petsalaki
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.
Anti-CLK2 antibody (ab86147) at 0.5 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 60 kDa Observed band size : 60 kDa Additional bands at : 36 kDa,64 kDa,94 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 8 minutesHuman Dual specificity protein kinase CLK2 contains a number of potential phosphorylation sites (SwissProt) which may explain the banding pattern at a higher molecular weight than predicted (64 kDa).
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