Detection of Neuropilin‑1 in Mouse CD4+ Splenocytes by Flow Cytometry.
Mouse CD4+ splenocytes were stained with Rabbit Anti-Human/Mouse FoxP3 Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC8214G) and either (A) Rat Anti-Mouse Neuropilin‑1 Alexa Fluor® 647‑conjugated Monoclonal Antibody (Catalog # FAB59941R) or (B) Rat IgG2A Alexa Fluor 647 Isotype Control (Catalog # IC006R). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
Neuropilin-1 (Nrp‑1, previously Neuropilin; also CD304) is a 130‑140 kDa type I transmembrane (TM) glycoprotein that regulates axon guidance and angiogenesis. The full-length 923 amino acid (aa) mouse Nrp‑1 contains a 835 aa extracellular domain (ECD) that shares 98% aa identity with rat and 93% aa identity with human, equine, bovine and canine Nrp-1. The ECD contains two N‑terminal CUB domains (termed a1a2), two domains with homology to coagulation factors V and VIII (b1b2) and a MAM (meprin) domain (c). The sema domains of Class III secreted semaphorins such as Sema3A bind Nrp‑1 a1a2. Heparin, the heparin-binding forms of VEGF (VEGF165, VEGF-B and VEGF-E), PlGF (PlGF2), and the C‑terminus of Sema3 bind the b1b2 region. Nrp‑1 and Nrp-2 share 48% aa identity within the ECD and can form homo‑ and hetero‑oligomers via interaction of their MAM domains. Neuropilins show partially overlapping expression in neuronal and endothelial cells during development. Both neuropilins act as co‑receptors with multiple molecules, which for Nrp-1 includes Sema3A through Sema3F, Plexin A1 through A4, Plexin B1 and D1. It also interacts with Robo1 and as noted, Nrp-2, as a heterodimer, which binds Sema3C. Finally, it has recently been found to bind miRNA that are complexed to AGO2 in the extracellular space. Both are co‑receptors with VEGF R2 (also called KDR or Flk‑1) for VEGF165 binding. Sema3A signaling can be blocked by VEGF165, which has higher affinity for Npn-1. Nrp-1 is preferentially expressed in developing or remodeling arteries. Nrp‑1 is also expressed on multiple cell types, including keratinocytes, Nrp-1+ T cells, Schwann cells, macrophages, vascular and lymphatic endothelium, breast duct epithelium, hepatic stellate cells, and neural crest cells that give rise to both sensory and autonomic ganglia.
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