Recombinant Human Neuropilin-1 Fc Avi-tag Protein, CF

Catalog #: AVI10455 Datasheet / COA / SDS

Biotinylated

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AVI10455-050

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Neuropilin-1 Fc Avi-tag Protein, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human VEGF165 (Catalog # 293-VE) is immobilized at 25.0 ng/mL (100 µL/well), Biotinylated Recombinant Human Neuropilin‑1 Fc Chimera Avi-tag binds with an ED₅₀ of 1.25-12.5 ng/mL. The biotin to protein ratio is greater than 0.7 as determined by the HABA assay.

Source

Human embryonic kidney cell, HEK293-derived human Neuropilin-1 protein

Human Neuropilin-1 (Phe22-Lys852) Accession # ABY87548.1	IEGRMD	Human IgG₁ (Pro100-Lys330)	Avi-tag
N-terminus			C-terminus

Accession #

ABY87548.1

N-terminal Sequence
Analysis

Phe22

Structure / Form

Disulfide-linked homodimer, biotinylated via Avi-tag

Predicted Molecular Mass

122 kDa

SDS-PAGE

120-140 kDa, under reducing conditions

Product Datasheets

AVI10455

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI10455

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Recombinant Human VEGF 165 (239-VE) is immobilized at 25.0 ng/mL (100 μL/well), Biotinylated Recombinant Human Neuropilin-1 Fc Chimera Avi-tag (Catalog # AVI10455) binds with an ED₅₀ of 1.25-12.5 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human Neuropilin-1 Fc Chimera Avi-tag Protein (Catalog # AVI10455) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 120-140 kDa and 240-280 kDa, respectively.

Reconstitution Calculator

Background: Neuropilin-1

Neuropilin-1 (NRP1), also known as CD304, is a type I transmembrane glycoprotein involved in embryonic development, axon guidance and vasculature formation (1‑4). The Neuropilin family is comprised of two isoforms, NRP1 and NRP2, that arose due to gene duplication and share 44% amino acid (aa) sequence identity (1). The extracellular domain (ECD) of Neuropilins are structurally similar containing two N‑terminal CUB domains (termed a1/a2), two domains with homology to coagulation factors V and VIII (b1/b2) and a MAM (meprin) domain (5). The ECD of NRP1 shares 93% aa sequence identity with mouse and rat NRP1. Several C‑terminally divergent splice variants of NRP1 and NRP2 lacking the MAM or transmembrane domain have been identified (5, 6). Neuropilins show partially overlapping expression in neuronal and endothelial cells during development (1, 2). They form homo- and hetero-oligomers via interaction of their MAM domains and share overlapping sets of ligands and functions (1, 7). Both Neuropilins act as co-receptors with Plexins and Semaphorins to help mediate axon repulsion (8, 9). NRP1 preferentially binds SEMA3A and PLXNA4, while NRP2 prefers binding to SEMA3F and PLXNA3 (8‑10). NRP1 is capable of binding VEGF and forming signaling complexes with VEGFR2 to mediate cell migration (10, 11). There is evidence that NRP1 with VEGF plays a role in stemness of breast cancer cell by activating Wnt/b-catenin pathway (12). Additionally, NRP1 is expressed on dendritic cells and mediates DC‑induced T cell proliferation (13). NRP1 is a marker of CD4+ Treg cells and a population of CD8+ T-cells infiltrating solid tumors and immunotherapies blocking NRP1 synergize with anti-PD1 to enhance CD8+ proliferation and response (14). NRP1 has been shown to bind the S1 fragment of the SARS-CoV-2 spike protein and may potentially provide a therapeutic target for COVID-19 (15). Our Avi-tag Biotinylated NRP1 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

Bielenberg, D.R. et al. (2006) Exp. Cell Res. 312:584.
Gu, C. et al. (2003) Dev. Cell 5:45.
He, Z. and M. Tessier-Lavigne (1997) Cell 90:739.
Soker, S. et al. (1998) Cell 92:735.
Nakamura, F. and Goshima, Y. (2002) Adv Exp Med Biol. 515:55.
Cackowski, F.C. et al. (2004) Genomics 84:82.
Guo, H.F. and Vander Kooi, C.W. (2015) J Biol Chem. 290:29120.
Yaron, A. et al. (2005) Neuron 45:513.
Schwarz, Q. and Ruhrberg, C. (2010) Cell Adh Migr. 4:61
Gu, C. et al. (2002) J. Biol. Chem. 277:18069
Herzog, B. et al. (2011) Mol Biol Cell. 22:2766).
Zhang, L. et al. (2017) Cell Physiol. Biochem. 44:1251.
Tordjman, R. et al. (2002) Nat. Immunol. 3:477.
Leclerc, M. et al. (2019) Nat Commun. 10:3345.
Daly, J. et al. (2020) Science. 6518:861.

Entrez Gene IDs

8829 (Human); 18186 (Mouse); 246331 (Rat); 100286859 (Porcine)

Alternate Names

BDCA4; BDCA-4; CD304 antigen; CD304; DKFZp686A03134; DKFZp781F1414; neuropilin 1; Neuropilin1; Neuropilin-1; NP1; NRP; NRP1; transmembrane receptor; Vascular endothelial cell growth factor 165 receptor; VEGF165R