Human Prostate Specific Antigen/KLK3 HEK293 Overexpression Lysate: Product Information
This Human Prostate Specific Antigen/KLK3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Prostate Specific Antigen/KLK3 protein (Cat: 10771-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human KLK3 (P07288) (Met 1-Pro 261) was fused with a polyhistidine tag at the C-terminus.
The recombinant human KLK3 consists of 255 amino acids and has a predicted molecular mass of 28.3 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh KLK3 is approximately 35 kDa.
Human Prostate Specific Antigen/KLK3 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 Prostate Specific Antigen/KLK3 HEK293 Overexpression Lysate: Alternative Names
Human APS Overexpression Lysate; Human Hk3 Overexpression Lysate; Human KLK2A1 Overexpression Lysate; Human PSA Overexpression Lysate
Prostate Specific Antigen/KLK3 Background Information
KLK3(Kallikrein 3), also known as Prostate Specific Antigen (PSA), kallikrein-related peptidase 3, Gamma-seminoprotein, is a secreted protein of the glandular kallikrein subfamily of serine proteases.KLK3 contains one peptidase S1 domain.KLK3 is a glycoprotein produced almost exclusively by the prostate gland. Growing evidence suggests that many kallikreins are implicated in carcinogenesis and some have potential as novel cancer and other disease biomarkers.
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