Human Syndecan-1/CD138 HEK293 Overexpression Lysate: Product Information
This Human Syndecan-1/CD138 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Syndecan-1/CD138 protein (Cat: 11429-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human SDC1 (NP_002988.3) extracellular domain (Met 1-Glu 251) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant human SDC1 consists of 245 amino acids and predictes a molecular mass of 25.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh SDC1 is approximately 48-55 kDa due to glycosylation.
Human Syndecan-1/CD138 HEK293 Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells 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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Human Syndecan-1/CD138 HEK293 Overexpression Lysate: Alternative Names
Human CD138 Overexpression Lysate; Human SDC Overexpression Lysate; Human SYND1 Overexpression Lysate; Human syndecan Overexpression Lysate; Human Syndecan-1 Overexpression Lysate
Syndecan-1/CD138 Background Information
Syndecan-1 also known as SDC1 and CD138, is the most extensively studied member of the syndecan family. It is found mainly in epithelial cells, but its expression is developmentally regulated during embryonic development. Syndecan-1/SDC1/CD138 has been shown to mediate cell adhesion to several ECM molecules, and to act as a coreceptor for fibroblast growth factors, potent angiogenic growth factors involved also in differentiation. Syndecan-1/SDC1/CD138 expression is reduced during malignant transformation of various epithelia, and this loss correlates with the histological differentiation grade of squamous cell carcinomas, lacking from poorly differentiated tumours. In squamous cell carcinomas of the head and neck, positive syndecan-1 expression correlates with a more favourable prognosis. Experimental studies on the role of Syndecan-1 in malignant transformation have shown that Syndecan-1/SDC1/CD138 expression is associated with the maintenance of epithelial morphology, anchorage-dependent growth and inhibition of invasiveness in vitro.
Inki P, et al. (1996) The role of syndecan-1 in malignancies. Ann Med. 28(1): 63-7.
Subramanian SV, et al. (1997) Regulated shedding of syndecan-1 and -4 ectodomains by thrombin and growth factor receptor activation. J Biol Chem. 272(23): 14713-20.
Park PW, et al. (2001) Exploitation of syndecan-1 shedding by Pseudomonas aeruginosa enhances virulence. Nature. 411(6833): 98-102.
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