Recombinant Human Nidogen-2 Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Nidogen-2 Protein, CF Summary

• Details of Functionality: Measured by the ability of the immobilized protein to support the adhesion of SVEC4‑10 mouse vascular endothelial cells. When 4 x 10⁴ cells/well are added to Recombinant Human Nidogen-2 coated plates (30 µg/mL with 100 µL/well), approximately 30-50% will adhere after one hour at 37° C.
  Optimal dilutions should be determined by each laboratory for each application.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Nidogen-2 protein
  Leu31-Lys1375 (Gly832Ala), with an N-terminal 9-His tag
• Accession #: Q14112
• N-terminal Sequence: His
• Protein/Peptide Type: Recombinant Proteins
• Gene: NID2
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 149.4 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 140-190 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Nidogen-2 Protein, CF

• NID2
• NID-2
• nidogen 2 (osteonidogen)
• Nidogen2
• Nidogen-2
• Osteonidogen

Background

Nidogen-2 (also named entactin-2) is a 200 kDa, secreted, monomeric basement membrane glycoprotein (1). Nidogens-1 and 2 are expressed in nearly all basement membranes (1-3) where they interact with laminins, collagen type IV and proteoglycan family members to form structural scaffolds (4, 5). In mouse, Nidogens 1 and 2 appear to substitute for each other. Deletion of one nidogen gives a mild phenotype, but deletion of both nidogens is lethal (6, 7). Affinity of laminin binding is much lower for human Nidogen-2 than that of mouse Nidogen-2, indicating that human Nidogen-2 may not be a strict substitute for Nidogen-1 (1). Both nidogens bind perlecan and collagens I and IV, but only Nidogen-1 binds fibulins (1, 3). The two nidogens show approximately 50% amino acid (aa) identity in human and are structurally similar (1, 4, 6). Cleavage of a 28 aa signal sequence from human Nidogen-2 produces a 1219 aa mature protein containing three globular domains
(G1-3) separated by a link region and an extended rod-shaped segment. The G1 domain is reported to bind type IV collagen, the G2 Nidogen ( beta -barrel) domain interacts with perlecan, and the C-terminal G3 beta -propeller structure is associated with laminin binding. The mucin-like link region is longer in Nidogen-2 than nidogen-1, and contains both N- and O-glycosylation (2, 8). There is one EGF-like motif and a short peptide that ligates alpha ₃ beta ₁ integrins. The rod-shaped segment contains four additional EGF-like motifs, two of which bind calcium, and two thyroglobulin type 1 domains that serve as a binding site for alpha _v beta ₃ integrins. Mature human Nidogen-2 is 80% aa identical to both mouse and rat Nidogen-2, and 73% aa identical to both canine and bovine Nidogen-2.

1. Kohfeldt, K. et al. (1998) J. Mol. Biol. 282:99.
2. Miosge, N. et al. (2001) Histochem. J. 33:523.
3. Salmivirta, K. et al. (2002) Exp. Cell Res. 279:188.
4. Hohenester, E. and J. Engel (2002) Matrix Biol. 21:115.
5. Charonis, A. et al. (2005) Curr. Med. Chem. 12:1495.
6. Schymeinsky, J. et al. (2002) Mol. Cell. Biol. 22:6820.
7. Bader, B.L. et al. (2005) Mol. Cell. Biol. 25:6846.

Publications for Nidogen-2 (3385-ND)(1)

Publication using 3385-ND

• Hendrickx AP, van Luit-Asbroek M, Schapendonk CM, van Wamel WJ, Braat JC, Wijnands LM, Bonten MJ, Willems RJ SgrA, a nidogen-binding LPXTG surface adhesin implicated in biofilm formation, and EcbA, a collagen binding MSCRAMM, are two novel adhesins of hospital-acquired Enterococcus faecium. Infect. Immun., 2009-09-08;77(11):5097-106. 2009-09-08 [PMID: 19737906] (Binding Assay, Bacteria)

Bioinformatics

• Gene Symbol: NID2
• Uniprot:
  • Q14112()