Detects mouse and rat Neuropilin-2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 25% cross‑reactivity with recombinant human Neuropilin-2 is observed, and less than 5% cross-reactivity with recombinant rat Neuropilin-1 is observed.
Polyclonal Goat IgG
Mouse myeloma cell line NS0-derived recombinant rat Neuropilin‑2 Gln23-Asp857 (Val809-Asp825 del) Accession # O35276
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Blockade of Receptor-ligand Interaction
In a functional ELISA, 1-3 µg/mL of this antibody will block 50% of the binding of 10 ng/mL of Recombinant Human VEGF165 (Catalog # 293-VE) to immobilized Recombinant Rat Neuropilin-2 Fc Chimera (Catalog # 567-N2) coated at 5 µg/mL (100 µL/well). At 60 μg/mL, this antibody will block >90% of the binding.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Mouse and Rat Neuropilin‑2 by Western Blot. Western blot shows lysates of C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse/Rat Neuropilin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Neuropilin‑2 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neuropilin‑2 in Rat Brain. Neuropilin‑2 was detected in perfusion fixed frozen sections of rat brain using Goat Anti-Mouse/Rat Neuropilin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Mouse and Rat Neuropilin‑2 by Simple WesternTM. Simple Western lane view shows lysates of C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line, loaded at 0.2 mg/mL. A specific band was detected for Neuropilin‑2 at approximately 140 kDa (as indicated) using 5 µg/mL of Goat Anti-Mouse/Rat Neuropilin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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 (Npn-1, previously known as Neuropilin) and Npn-2 (previously known as Npn-1-related molecule) are type I transmembrane proteins that bind members of the class III secreted semaphorin subfamily which are implicated in repulsive axon guidance. The extracellular domain of these proteins is composed of two N-terminal CUB (complement-binding) domains (domains a1 and a2), two domains with homology to coagulation factors V and VIII (domains b1 and b2) and a MAM (meprin) domain (domain c). In the absence of ligands, neuropilins can form homo- and hetero-oligomers via homophilic interactions of their MAM domains. At the amino acid sequence level, Npn-1and Npn-2 share 44% identity. Npn-1 and Npn-2 show different binding specificities for different members of the semaphorin family. The expression patterns of Npn-1 and Npn-2 in developing neurons of the central and peripheral nervous systems are largely, though not completely nonoverlapping. Npn‑1 and Npn-2 are also expressed by endothelial and tumor cells and have been shown to be isoform-specific receptors for VEGF165. Npn‑1 was also reported to bind PlGF-2 and the VEGF-like protein from of virus NZ2.
Fujisawa, H. and T. Kitsukawa (1998) Curr. Opin. Neurobiol. 8:587.
Neufeld, G. et al. (1999) FASEB J. 13:9.
Poltorak, Z. et al. (2000) J. Biol. Chem. 275:18040.
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species: Mouse.We have 2 reviews tested in 2 applications: Immunohistochemistry, Whole mount immunofluorescence.