The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
1/10 - 1/50.
One family of inhibitory axon guidance molecules is the semaphorins. The semaphorins include secreted, transmembrane, and GPI anchored extracellular molecules that are involved in regulating axon guidance by inhibiting axons from growing toward incorrect targets. Semaphorin 3A (Sema3A) may play a particularly interesting role in limiting axon regeneration since it is expressed in meningeal fibroblasts that invade the injured spinal cord and surround the glial scar. In addition, the Sema3A co receptors, Neuropilin 1 and Plexin A1, are expressed on axons that regenerate up to the injured region, but do not cross this Sema3A containing region. Thus, Sema3A and its co receptors may have important roles in regulating axon guidance during neuronal development and after neuronal injury.
ab80011, at a 1/10 dilution, staining Semaphorin 3A in formalin fixed, paraffin embedded human breast carcinoma tissue by Immunohistochemistry. Peroxidase conjugated secondary antibody was used, followed by DAB staining.
ICC/IF image of ab80011 stained MCF7 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 (ab80011, 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.