膜杰作
Star Staining
分子别名（Synonym）
SPAM1,PH-20,HYAL3,HYA1,HYAL1,HYAL5,SPAG15
表达区间及表达系统（Source）
Human PH20, Tag Free (PH0-H5219) is expressed from human 293 cells (HEK293). It contains AA Leu 36 - Ser 490 (Accession # P38567-1).
Predicted N-terminus: Leu 36
Request for sequence
蛋白结构（Molecular Characterization）
This protein carries no "tag"
The protein has a calculated MW of 52.6 kDa. The protein migrates as 65-70 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
内毒素（Endotoxin）
Less than 1.0 EU per μg by the LAL method.
纯度（Purity）
>95% as determined by SDS-PAGE.
>95% as determined by SEC-MALS.
制剂（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.
重构方法（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.
存储（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:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
电泳（SDS-PAGE）
Human PH20, Tag Free on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
SEC-MALS
The purity of Human PH20, Tag Free (Cat. No. PH0-H5219) is more than 95% and the molecular weight of this protein is around 53-71 kDa verified by SEC-MALS.
Report
活性（Bioactivity）
The activity of PH20 is measured by its ability to hydrolyze HA in turbidimetric assay (45 minute assay). The specific activity is >55000 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).
Protocol
背景（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.
