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.
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.
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: 使用指南
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).
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.
Human PRSS3 HEK293 Overexpression Lysate: 别称
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
Trypsin-3, also known as Trypsin III, brain trypsinogen, Serine protease 3 and PRSS3, is a secreted protein that belongs to the peptidase S1 family. Trypsin-3 / PRSS3 is expressed is in pancreas and brain. It contains one peptidase 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.