Human C-Reactive Protein HEK293 Overexpression Lysate: Product Information
This Human C-Reactive Protein overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of C-Reactive Protein protein (Cat: 11250-HNAH) from the overexpression lysate was verified.
A DNA sequence encoding the human CRP (P02741-1) (Met1-Pro224) was expressed and purified.
The recombinant human CRP consists of 206 amino acids and has a predicted molecular mass of 23 kDa. As a result of glycosylation, the apparent molecular mass of CRP is approximately 26 kDa in SDS-PAGE under reducing conditions.
Human C-Reactive Protein 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 C-Reactive Protein HEK293 Overexpression Lysate: Alternative Names
Human C-Reactive Protein Overexpression Lysate; Human PTX1 Overexpression Lysate
C-Reactive Protein Background Information
C-reactive protein (CRP) is synthesized by the liver in response to factors released by fat cells. It is a member of the pentraxin family of proteins. The levels of CRP rise in response to inflammation. Human C-reactive protein (CRP) is the classical acute phase reactant, the circulating concentration of which rises rapidly and extensively in a cytokine-mediated response to tissue injury, infection and inflammation. Serum CRP values are routinely measured, empirically, to detect and monitor many human diseases. However, CRP is likely to have important host defence, scavenging and metabolic functions through its capacity for calcium-dependent binding to exogenous and autologous molecules containing phosphocholine (PC) and then activating the classical complement pathway. CRP may also have pathogenic effects and the recent discovery of a prognostic association between increased CRP production and coronary atherothrombotic events is of particular interest.
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