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Human PH20 / SPAM1 Protein, His Tag (active enzyme) DMF

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  • Synonym
    SPAM1,PH-20,HYAL3,HYA1,HYAL1,HYAL5,SPAG15
  • Source
    Human PH20, His Tag(PH0-H5225) is expressed from human 293 cells (HEK293). It contains AA Leu 36 - Tyr 482 (Accession # P38567-1).
    Predicted N-terminus: Leu 36
  • Molecular Characterization
    PH20 Structure

    This protein carries a polyhistidine tag at the C-terminus.

    The protein has a calculated MW of 53.0 kDa. The protein migrates as 60-66 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM NaCl, pH7.5 with trehalose as protectant.

    Contact us for customized product form or formulation.

  • Reconstitution

    Please see Certificate of Analysis for specific instructions.

    For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

  • Storage

    For long term storage, the product should be stored at lyophilized state at -20°C or lower.

    Please avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 12 months under sterile conditions after reconstitution.
SDS-PAGE
PH20 SDS-PAGE

Human PH20, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity

The activity of Human PH20, His Tag (Cat. No. PH0-H5225) is measured by its ability to hydrolyze HA in turbidimetric assay (45 minute assay). The specific activity is >60,000 U/mg. (Unit Definition: One unit of Hyaluronidase activity will cause a change in A600 of 0.330 per minute at pH5.35 at 37 °C  in a 2.0 mL reaction mixture) (QC tested).

  • Background
    Hyaluronidase PH-20 is also known as Sperm adhesion molecule 1 (SPAM1) and Sperm surface protein PH-20, which belongs to the glycosyl hydrolase 56 family, SPAM1 / PH-20 is expressed in testis. SPAM-1 / PH20 random hydrolysis of (1->4)-linkages between N – acetyl – beta – D – glucosamine and D-glucuronate residues in hyaluronate. SPAM-1 / PH20 involved in sperm-egg adhesion. Upon fertilization sperm must first penetrate a layer of cumulus cells that surrounds the egg before reaching the zona pellucida. The cumulus cells are embedded in a matrix containing hyaluronic acid which is formed prior to ovulation. SPAM1 aids in penetrating the layer of cumulus cells by digesting hyaluronic acid.
  • Clinical and Translational Updates

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