Product Specifications for Recombinant Human Nidogen-2 Protein, CF
Chinese Hamster Ovary cell line, CHO-derived human Nidogen-2 protein Leu31-Lys1375 (Gly832Ala), with an N-terminal 9-His tag
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
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.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
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
Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitute at 100 μg/mL in sterile PBS.
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.
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.
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.
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