Human Prostasin/PRSS8 HEK293 Overexpression Lysate: Product Information
This Human Prostasin/PRSS8 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Prostasin/PRSS8 protein (Cat: 10287-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human Prss8 (NP_002764.1) (Ala 30-Arg 322) was fused with a signal peptide at the N-terminus and a polyhistidine-tag at the C-terminus.
The mature recombinant human Prss8 consists of 304 amino acids and predicts a molecular mass of 32.8 kDa. By SDS-PAGE under reducing conditions, the apparent molecular mass of rhPrss8 is approximately 40 kDa due to glycosylation.
Human Prostasin/PRSS8 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 Prostasin/PRSS8 HEK293 Overexpression Lysate: Alternative Names
Human CAP1 Overexpression Lysate; Human PROSTASIN Overexpression Lysate
Prostasin/PRSS8 Background Information
Prostasin (Prss8), also known as channel activating protease 1 (CAP1), is a trypsinlike serine peptidase, and plays important roles in epithelial physiology. It is originally purified as an active, soluble enzyme from human seminal fluid and is highly expressed in prostate, lung, kidney, salivary gland and pancreas. Prostasin is expressed as a glycosyl-phosphatidylinositol (GPI)-anchored membrane protein in prostate epithelial cells, and also exists as a secreted proteolytic enzyme possibly via tryptic cleavage of its COOH-terminal hydrophobic domain. Prostasin is found to activate the epithelial sodium channel (ENaC) which is tightly regulated and is critical for maintaining salt and fluid balance in the lung and kidney in both normal and pathological conditions. Accordingly, prostasin has been proposed as a target for therapeutic inhibition in cystic fibrosis. In addition, prostasin inhibits prostate and breast cancer cell invasion in vitro, suggesting a functional role as a suppressor of tumor invasion, as well as a regulator of gene expression during inflammation.
protease, serine, 8
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Martin J., et al.,(2004), The sequence and analysis of duplication-rich human chromosome 16.Nature 432:988-994.
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