Recombinant Human Nidogen-2 Protein, CF

Catalog #: 3385-ND
1 Citation Datasheet / COA / SDS

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3385-ND-050

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

Recombinant Human Nidogen-2 Protein, CF Summary

Product Specifications

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

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
Analysis

His

Predicted Molecular Mass

149.4 kDa

SDS-PAGE

140-190 kDa, reducing conditions

Product Datasheets

3385-ND

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.

3385-ND

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in sterile 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.

Reconstitution Calculator

Background: Nidogen-2

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.

References

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

Entrez Gene IDs

22795 (Human); 18074 (Mouse); 302248 (Rat)

Alternate Names

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

Related Research Areas

Citation for Recombinant Human Nidogen-2 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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