Human Protein C HEK293 Overexpression Lysate

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Human Protein C HEK293 Overexpression Lysate: Product Information

Product Description
This Human Protein C overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Protein C protein (Cat: 10196-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human PROC (P04070) (Met1-Pro461) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human PROC comprises 454 amino acids and has a predicted molecular mass of 51.5 kDa. The apparent molecular mass of the protein is approximately 56-64 and 45 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human Protein C 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.

Human Protein C HEK293 Overexpression Lysate: Alternative Names

Human APC Overexpression Lysate; Human PC Overexpression Lysate; Human PROC Overexpression Lysate; Human PROC1 Overexpression Lysate; Human THPH3 Overexpression Lysate; Human THPH4 Overexpression Lysate

Protein C Background Information

PROC (protein C gene) is a 62-kDa vitamin K-dependent plasma zymogen which, after activation to serine protease, plays an important role in the physiologic regulation of blood coagulation. Originally thought to be synthesized exclusively by the liver, recent reports have shown that PROC is also produced by many other cells including pancreatic islet β cells. Protein C a vitamin K-dependent plasma glycoprotein and is one of the body’s natural anticoagulants circulating at 7 nmol/L as an inactive protease zymogen.
Full Name
protein C, inactivator of coagulation factors Va and VIIIa
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