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 42 kDa (predicted molecular weight: 16 kDa).
Use a concentration of 5 µg/ml.
Has heparin binding activity, and growth promoting activity. Involved in neointima formation after arterial injury, possibly by mediating leukocyte recruitment. Also involved in early fetal adrenal gland development.
The observed molecular weight of 42 kDa is the combined molecular weight of the full-length recombinant Midkine protein (~140 amino-acids) and the tag.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Midkine antibody (ab36038)This image is courtesy of an abreview submitted by Sophie Pezet, ESPCI, France
IHC-FoFr image of Midkine (ab36038) staining on Rat Spinal cord sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the cords were cryoprotected in sucrose 30% overnight. Tissues were then cut using a cryostat and the immunostainings were preformed using the ‘free floating’ technique.
Image B is a high power magnification of Image A.
ICC/IF image of ab36038 stained DU145 cells. The cells were 4% formaldehyde 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 (ab36038, 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.