Recombinant Bovine SPAM1 Protein, CF
R&D Systems, part of Bio-Techne | Catalog # 6436-GH
Key Product Details
Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived bovine SPAM1 protein
Met1-Thr497, with a C-terminal 6-His tag
Met1-Thr497, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Leu36
Predicted Molecular Mass
53 kDa
SDS-PAGE
60-70 kDa, reducing conditions
Activity
Measured by its ability to hydrolyze S-35 labeled bovine cartilage chondroitin sulfate.
<20 ng of recombinant bovine SPAM1 is required for the 50% hydrolysis of 10 μg chondroitin sulfate, as measured under the described conditions.
<20 ng of recombinant bovine SPAM1 is required for the 50% hydrolysis of 10 μg chondroitin sulfate, as measured under the described conditions.
Formulation, Preparation and Storage
6436-GH
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: SPAM1
GPI‑anchored enzyme located on the sperm surface and inner acrosomal membrane (4). SPAM1 degrades hyaluronic acid (HA), a major structural glycosaminoglycan found in extracellular matrices and basement membranes. The enzyme activity enables sperm to penetrate through the HA-rich cumulus cell layer surrounding the oocyte and therefore facilitates the fertilization process (5). However, detailed enzymatic analysis of this enzyme is hindered by the limited techniques/methods available to monitor the HA degradation products (6). A novel method for analyzing SPAM1 activity was utilized here. Because of the structural similarity between HA (repeating units of GlcA beta1-3GlcNAc) and chondroitin sulfate (repeating units of GlcA beta1-3GalNAc), the enzyme is also able to hydrolyze chondroitin sulfate. In this assay, radiolabeled chondroitin sulfate was digested with recombinant SPAM1. Degradation products were then separated using a polyacrylamide electrophoresis and visualized with an X-ray film (7). The bovine SPAM1 is 63% identical to human homologue in sequence.
References
- Lathrop, W.F. et al. (1990) J. Cell Biol. 111:2939.
- Lin, Y. et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90:10071
- Csoka, A.B. et al. (2001) Matrix Biol. 20:499.
- Phelps, B.M. et al (1988) Science 240:4860.
- Gmachl, M. et al. (1993) FEBS Lett. 336:545.
- Hofinger, E.S et al. (2007) Glycobiology 17:963.
- Wu, Z.L. et al. (2010) BMC Biotechnol. 10:11.
Long Name
Sperm Adhesion Molecule 1
Alternate Names
HYA1, HYAL3, HYAL5, PH20, SPAG15
Gene Symbol
SPAM1
UniProt
Additional SPAM1 Products
Product Documents for Recombinant Bovine SPAM1 Protein, CF
Product Specific Notices for Recombinant Bovine SPAM1 Protein, CF
For research use only
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