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.
Ab23393 staining human normal ileum. Staining is localized to the cytoplasm. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Western blot - Anti-Semaphorin 3A antibody (ab23393)
Immunohistochemistry (Frozen sections) - Anti-Semaphorin 3A antibody (ab23393)Image courtesy of Dr Ruxandra Sirbulescu by Abreview.
ab23393 staining Semaphorin 3A in 16 µm thick sections of Apteronotus leptorhynchus brain, spinal cord by Immunohistochemistry (Frozen sections). Tissue was fixed in 2% paraformaldehyde, permeabilized using 0.3% Triton X-100, then blocked with 3% sheep serum, 1% BSA, 1% teleostean gelatine in TBS for 1 hour at 24°C and then incubated with ab23393 at a 1/50 dilution for 18 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/200 dilution.
Rienks M et al. Sema3A promotes the resolution of cardiac inflammation after myocardial infarction. Basic Res Cardiol112:42 (2017).
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Inoue-Torii A et al. The level of urinary semaphorin3A is associated with disease activity in patients with minimal change nephrotic syndrome. Int J Nephrol Renovasc Dis10:167-174 (2017).
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