Human Neuropilin-1 EZ-Set™ ELISA Kit (DIY Antibody Pairs)

NRP1 (Neuropilin 1) also known as NP1, NRP, BDCA4 or VEGF165R, is a membrane-bound coreceptor to a tyrosine kinase receptor for both vascular endothelial growth factor (VEGF) and semaphorin family members. NRP1 plays versatile roles in angiogenesis, axon guidance, cell survival, migration, and invasion. By somatic cell hybrid analysis, the NRP1 gene was mapped to chromosome 10. NRP1 bounds PGF1 with lower affinity. NRP1-mediated interactions are a necessary element in the initiation of the primary immune response and offer another example, like that of agrin, of a molecule shared by neurologic and immunologic synapses. After T-cell contact with DC, T-cell NRP1 colocalized with CD3 in the immunologic synapse and, sometimes, also at the opposite pole of the T cell. Soluble NRP1 interacts in a homophilic fashion with NRP1 on both DC and T cells, and this binding can be inhibited by blocking antibodies to NRP1. Furthermore, selective NRP1 inhibition in this model suppressed neovascular formation substantially.

The expression of isoforms 1 and 2 does not seem to overlap. Isoform 1 is expressed by the blood vessels of different tissues. In the developing embryo it is found predominantly in the nervous system. In adult tissues, it is highly expressed in heart and placenta; moderately in lung, liver, skeletal muscle, kidney and pancreas; and low in adult brain. Isoform 2 is found in liver hepatocytes, kidney distal and proximal tubules.

Adaptive Immunity, Angiogenesis, Axonal Guidance Proteins, Cardiovascular, Growth and Development, Growth Factors, Immunology, Neurology Process, Neuroscience

The membrane-bound isoform 1 is a receptor involved in the development of the cardiovascular system, in angiogenesis, in the formation of certain neuronal circuits and in organogenesis outside the nervous system. It mediates the chemorepulsant activity of semaphorins. It binds to semaphorin 3A, The PLGF-2 isoform of PGF, The VEGF-165 isoform of VEGF and VEGF-B. Coexpression with KDR results in increased VEGF-165 binding to KDR as well as increased chemotaxis. It may regulate VEGF-induced angiogenesis.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1353-CA Goat anti-human Neuropilin-1 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1353-DA Biotinylated goat anti-human Neuropilin-1 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1353-ST Lyophilized recombinant human Neuropilin-1 standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1353-CA Goat anti-human Neuropilin-1 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1353-DA Biotinylated goat anti-human Neuropilin-1 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1353-ST Lyophilized recombinant human Neuropilin-1 standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1353-CA Goat anti-human Neuropilin-1 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1353-CA Goat anti-human Neuropilin-1 polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1353-DA Biotinylated goat anti-human Neuropilin-1 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ1353-DA Biotinylated goat anti-human Neuropilin-1 polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1353-ST Lyophilized recombinant human Neuropilin-1 standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ1353-ST Lyophilized recombinant human Neuropilin-1 standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.016 0.137 0.269 0.476 0.846 1.395 1.808 2.043 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.016 0.137 0.269 0.476 0.846 1.395 1.808 2.043 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.016 0.137 0.269 0.476 0.846 1.395 1.808 2.043 O.D. 0.016 0.137 0.269 0.476 0.846 1.395 1.808 2.043

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)