Recombinant Mouse Nidogen-2 Protein, CF

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

Recombinant Mouse 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 5 x 10⁴ cells/well are added to Recombinant Mouse Nidogen-2 coated plates (10 µg/mL with 100 µL/well), >40% will adhere after one hour at 37 °C.
  Optimal dilutions should be determined by each laboratory for each application.
• Source: Mouse myeloma cell line, NS0-derived mouse Nidogen-2 protein
  Leu31-Lys1403 with an N-terminal 6-His tag
• Accession #: NP_032721
• N-terminal Sequence: Not obtained, predicted His-tag
• 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: 151.5 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: 170-185 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 PBS.

Notes

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

Alternate Names for Recombinant Mouse 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). Mice deleted for both nidogen genes die at birth due to delayed basement membrane maturation of the lungs and heart (7). However, deletion of one nidogen gives a mild phenotype, indicating that their functions overlap (6, 7). In adult, muscle Nidogen-2 expression is concentrated in the basal lamina of synapses and is required for their maturation and maintenance (8). Both nidogens bind laminin, perlecan and collagens I and IV, but only Nidogen-1 binds fibulins (1, 3). The two nidogens share approximately 40% amino acid (aa) identity in mouse and are structurally similar (1, 4, 6). Cleavage of a 30 aa signal sequence from mouse Nidogen-2 produces a 1373 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). It contains one EGF-like motif, and an RGD motif that binds 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. If species‑specific insertions and deletions are disregarded, mature mouse Nidogen-2 shares 80%, 92%, 74%, 73%, 71% and 68% aa identity with human, rat, bovine, porcine, canine and equine Nidogen-2, respectively.

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.
8. Fox, M.A. et al. (2008) Neural Dev. 3:24.

Publications for Nidogen-2 (6760-ND)(1)

Publication using 6760-ND

• Patlaka C, Mai H, Lang P, Andersson G The growth factor-like adipokine tartrate-resistant acid phosphatase 5a interacts with the rod G3 domain of adipocyte-produced nidogen-2. Biochem Biophys Res Commun, 2014-10-29;454(3):446-452. 2014-10-29 [PMID: 25450682] (Bioassay, Western Blot)

Bioinformatics

• Gene Symbol: Nid2
• Uniprot:
  • NP_032721()