Detection of Neuropilin‑1 in Mouse Splenocytes by Flow Cytometry.
Mouse splenocytes were stained with Rat Anti-Mouse CD4 APC-conjugated Monoclonal Antibody (Catalog # FAB554A) and either (A) Rat Anti-Mouse Neuropilin‑1 Monoclonal Antibody (Catalog # MAB59941) or (B) Rat IgG2A Isotype Control (Catalog # MAB006) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B).
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Neuropilin-1 (Nrp‑1, previously neuropilin; also CD304) is a 130‑140 kDa type I transmembrane (TM) glycoprotein that regulates axon guidance and angiogenesis (1‑4). The full-length 923 amino acid (aa) mouse Nrp‑1 contains a 623 aa extracellular domain (ECD) that shares 98% aa identity with rat and 93% aa identity with human, equine, bovine and canine Nrp-1 (3, 4). 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). At least one splice variant that diverges at aa 587 and lacks the TM domain has been sequenced (5). This form is potentially a soluble antagonist, based on results from human Nrp‑1 splice variants (1, 6‑8). The sema domains of Class III secreted semaphorins such as Sema3A bind Nrp‑1 a1a2 (9). 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 (9, 10). 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 (1). Neuropilins show partially overlapping expression in neuronal and endothelial cells during development (1, 2). Both neuropilins act as co‑receptors with plexins, mainly plexin A3 and A4, to bind class III semaphorins that mediate axon repulsion (11). However, only Nrp‑1 binds Sema3A, and only Nrp‑2 binds Sema3F (1). Both are co‑receptors with VEGF R2 (also called KDR or Flk‑1) for VEGF165 binding (1). Sema3A signaling can be blocked by VEGF165, which has higher affinity for Npn-1 (12). Nrp-1 is preferentially expressed in developing or remodeling arteries (1, 2). Nrp‑1 is also expressed on dendritic cells and mediates DC‑induced T cell proliferation (13).
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