This Mouse CD63 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD63 protein (Cat: 50557-MNAH) from the overexpression lysate was verified.
A DNA sequence encoding the mouse CD63 (NP_031679.1) (Ala103-Ile203) was expressed.
The recombinant mouse CD63 consists of 101 amino acids and predicts a molecular mass of 11.5 kDa.
Mouse CD63 HEK293 Overexpression Lysate: 使用指南
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
The cluster of differentiation (CD) system is commonly used as cell markers in Immunophenotyping. Different kinds of cells in the immune system can be identified through the surface CD molecules which associating with the immune function of the cell. There are more than 320 CD unique clusters and subclusters have been identified. Some of the CD molecules serve as receptors or ligands important to the cell through initiating a signal cascade which then alter the behavior of the cell. Some CD proteins do not take part in cell signal process but have other functions such as cell adhesion. Cluster of differentiation 63 (CD63) is a member of the CD family and the transmembrane 4 superfamily, also known as the tetraspanin family. CD63 is a cell surface glycoprotein characterized by the presence of four hydrophobic domains. CD63 had functions in mediating signal transduction processes and then regulate a variety of cellular processes such as cell proliferation, activation and motility. It has been reported that CD63 protein associated with tumor progression and served as a blood platelet activation marker and the deficiency of this protein may be associated with Hermansky-Pudlak syndrome.
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