Sema3A Recombinant Protein

Growth cone collapse testing was carried out to verify recombinant Sema3A protein induces collapse. Briefly, dorsal root ganglion cells were plated and allowed to grow overnight in differentiation media. The next day the media was replaced with differentiation media or differentiation media with 1µg/ml of Sema3A protein. The cells were then incubated for 30 minutes, fixed, and stained with FITC conjugated antibodies specific for phalloidin (Part No. 90228). Representative images of each assay condition are shown below:

The purity of the Sema3A protein was also determined by SDS-PAGE. Purified protein (2µg) was run using non-denaturing conditions and then coomassie blue stained. The protein detected was approximately 125kD in size.

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Semaphorins are a large family of well conserved proteins that participate in a variety of cell signaling functions. However, they are best known for their role in development and regeneration within the nervous system. These affects have been shown to be primarily mediated by the class 3 semaporins (Sema3). In particular, the Sema3A family member acts as both a chemorepellant for axonal growth and a chemoattractant for dendritic processes1,2.

Here we provide recombinant Sema3A protein to facilitate researchers studying neurite outgrowth. The protein produced is an Fc-chimera containing the highly conserved semphorin domain, the Ig-like domain, and the highly positively charged C-terminal region (without the 20 amino acids at the C-terminus)3.

40µl of 0.25mg/ml recombinant protein.

Protein A affinity purified recombinant Sema3A protein in PBS buffer.

Store at -20°C or at -80°C for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.