This antibody gave a positive signal in the following lysates: Human Breast Tissue; Mouse Lung Tissue; Mouse Brain Tissue; Rat Brain Tissue; MCF7 Whole Cell; HepG2 Whole Cell; HeLa Whole Cell; Jurkat Whole Cell; A549, Human Glioblastoma Whole Cell.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
Use a concentration of 5 µg/ml.
RelevanceThe dominant pathway for biosynthesis of phosphatidylcholine occurs through the CDP-choline pathway. Choline kinase alpha (CHK) is the first enzyme in the pathway and may thereby play an upstream regulatory role in lipid transport and metabolism. CHK additionally catalyzes phosphorylation of ethanolamine.
ICC/IF image of ab88053 stained HeLa 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 (ab88053, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit 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.
Western blot - Anti-Choline kinase alpha antibody (ab88053)
All lanes : Anti-Choline kinase alpha antibody (ab88053) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate Lane 2 : Mouse lung normal tissue lysate - total protein (ab29297) Lane 3 : Brain (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 52 kDa Observed band size : 52 kDa Additional bands at : 72 kDa,78 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 1 minute
Immunocytochemistry/ Immunofluorescence - Anti-Choline kinase alpha antibody (ab88053)This image is courtesy of an anonymous abreview.
Immunocytochemistry/ Immunofluorescence analysis of human glioblastoma cells labeling Choline kinase alpha with ab88053 at 1/250 dilution. Cells were fixed in paraformaldehyde and permeabilized with TritonX-100. A goat anti-rabbit Alexa Fluor594 was used as the secondary antibody at 1/1000 dilution.