Human PRSS3 HEK293 Overexpression Lysate
Cat: 11866-H08HL
Cat: 11866-H08HL
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Human PRSS3 HEK293 Overexpression Lysate: Product Information
Product Description
This Human PRSS3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PRSS3 protein (Cat: 11866-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human PRSS3 isoform c (P35030-3) (Met 1-Ser 247) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The secreted recombinant pro form of the human PRSS3 consists of 243 amino acids and predictes a molecular mass of 26.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhPRSS3 is approximately 33 kDa due to glycosylation.
Human PRSS3 HEK293 Overexpression Lysate: Usage Guide
Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Optimal dilutions/concentrations should be determined by the end user.
Human PRSS3 HEK293 Overexpression Lysate: Alternative Names
Human MTG Overexpression Lysate; Human PRSS4 Overexpression Lysate; Human RP11-176F3.3 Overexpression Lysate; Human T9 Overexpression Lysate; Human TRY3 Overexpression Lysate; Human TRY4 Overexpression Lysate
PRSS3 Background Information
Trypsin-3, also known as Trypsin III, brain trypsinogen, Serine protease 3 and PRSS3, is a secreted protein which belongs to thepeptidase S1 family. Trypsin-3 / PRSS3 is expressed is in pancreas and brain. It contains onepeptidase S1 domain. Trypsin-3 / PRSS3 can degrade intrapancreatic trypsin inhibitors that protect against CP. Genetic variants that cause higher mesotrypsin activity might increase the risk for chronic pancreatitis (CP). A sustained imbalance of pancreatic proteases and their inhibitors seems to be important for the development of CP. The trypsin inhibitor-degrading activity qualified PRSS3 as a candidate for a novel CP susceptibility gene. Trypsin-3 / PRSS3 has been implicated as a putative tumor suppressor gene due to its loss of expression, which is correlated with promoter hypermethylation, in esophageal squamous cell carcinoma and gastric adenocarcinoma.
Full Name
protease, serine 3
References
- Venter JC. et al., 2001, Science 291:1304-51.
- Marsit,CJ. et al., 2005, Mol Carcinog 44 (2):146-50.
- Rowen, L. et al., 2005, Mol Biol Evol. 22 (8):1712-20.
- Rosendahl, J. et al., 2010, Pancreatology. 10 (2-3):243-9
