Product Specifications for Recombinant Mouse DSPG3 Protein, CF
Mouse myeloma cell line, NS0-derived mouse DSPG3 protein Ala20-Ile322, with a C-terminal 10-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
52-57 kDa, reducing conditions
Measured by the ability of the immobilized protein to inhibit the adhesion of Saos‑2 human osteosarcoma cells to human Fibronectin.
When 5 x 104 cells/well are added to Mouse DSPG3 and Human Fibronectin (0.5 μg/mL, Catalog # 1918-FN) coated plates, the cell adhesion is enhanced in a dose dependent manner. The ED50 for this effect is 0.3-1.2 μg/mL.
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 200 μg/mL in 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.
DSPG3 (dermatan sulfate proteoglycan 3), also called epiphycan (gene name EPYC) or PG-Lb, is a 95 ‑ 100 kDa class III subfamily member of the SLRP (small leucine-rich proteoglycans) family of molecules (1 ‑ 4). It is synthesized as a 322 amino acid (aa) precursor that contains a 19 aa signal sequence and a 303 aa mature region. The mature molecule contains three N-terminal O-glycosylation sites (including one found to bind dermatin sulfate) and a glutamic acid-rich motif followed by six leucine-rich repeats (LRR) that contain two potential N-glycosylation sites (2, 3). The LRR are flanked by two intrachain disulfide bonds (5). Class III SLRPs, including DSPG3, Osteoglycin and Opticin, contain 6 ‑ 7 LRR, while other classes contain 8 ‑ 10 LRR (1, 4). Collagen binding maps to the LRR (1). Mature mouse DSPG3 shares 97%, 85%, 84%, and 83% aa sequence identity with rat, human, canine and bovine DSPG3, respectively. DSPG3 is found predominantly in epiphyseal cartilage, although DSPG3 mRNA is also detected in mouse testis and human ligament and placenta (2‑4). In mouse, expression begins in mid-gestation as an intermediate marker for chondrogenesis, peaks at birth and declines thereafter (6, 7). Deletion of DSPG3 in mouse results in a mild postnatal phenotype that worsens synergistically when the co‑expressed SLRP biglycan is also deleted (7). The double-mutant phenotype includes premature osteoarthritis, indicating a role for DSPG3, in cooperation with other SLRPs, in stability of the collagen matrix and maintenance of joint integrity (7).
Kalamajski, S. and A. Oldberg (2010) Matrix Biol. 29:248.
Kurita, K. et al. (1996) Biochem. J. 318:909.
Deere, M. et al. (1996) Genomics 38:399.
Deere, M. et al. (1999) Genome Res. 9:449.
Swiss-Prot Accession # Q96645 and P70186
Johnson, J. et al. (1999) Dev. Dyn. 216:499.
Nuka, S. et al. (2010) Osteoarthritis Cartilage 18:88.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
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