The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).Can be blocked with Rat Gliomedin peptide (ab25746).
Use a concentration of 10 µg/ml.
Gliomedin is a member of the collagen superfamily, it is a glial ligand for neurofascin and NrCAM, two axonal immunoglobulin cell adhesion molecules that are associated with Na+ channels at the nodes of Ranvier. Gliomedin provides a glial cue for the formation of peripheral nodes of Ranvier. Gliomedin is expressed by myelinating Schwann cells and accumulates at the edges of each myelin segment during development, where it aligns with the forming nodes of ranvier. Eliminating the expression of gliomedin or the addition of a soluble extracellular domain of neurofascin to myelinating cultures abolishes node formation. Gliomedin is expressed in the PNS nodes of ranvier, but not in the CNS nodes of ranvier.
Gliomedin also displays high expression in murine and human hepatocellular carcinomas (HCC). Its restricted expression in normal tissues and unique early upregulation during tumor development make it an excellent candidate as a new clinical marker of HCC.
ab24483 detects a band of 70 kDa in Western Blot. Gliomedin contains a number of N-glycosylation sites (SwissProt data) which may explain the migration at a higher molecular weight than predicted based on primary sequence.
ICC/IF image of ab24483 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab24483, 10µg/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.