Human KLK4 HEK293 Overexpression Lysate

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Human KLK4 HEK293 Overexpression Lysate: Product Information

Product Description
This Human KLK4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of KLK4 protein (Cat: 11857-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human KLK4 (NP_004908.3) (Met 1-Ser 254) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The secreted recombinant human KLK4 consists of 239 amino acids and predictes a molecular mass of 25.8 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhKLK4 is approximately 30-35 kDa due to glycosylation.

Human KLK4 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 KLK4 HEK293 Overexpression Lysate: Alternative Names

Human AI2A1 Overexpression Lysate; Human ARM1 Overexpression Lysate; Human EMSP Overexpression Lysate; Human EMSP1 Overexpression Lysate; Human kallikrein Overexpression Lysate; Human KLK-L1 Overexpression Lysate; Human PRSS17 Overexpression Lysate; Human PSTS Overexpression Lysate

KLK4 Background Information

Kallikrein-4, also known as Enamel matrix serine proteinase 1, Kallikrein-like protein 1, KLK-L1, Serine protease 17, KLK4, PRSS17 and EMSP1, is a secreted protein which belongs to thepeptidase S1 family and Kallikrein subfamily. Kallikrein-4 / KLK4 is a serine protease expressed during enamel maturation, and proteolytic processing of the enamel matrix by KLK4 is critical for proper enamel formation. Kallikrein-4 / KLK4 contains onepeptidase S1 domain. Kallikrein-4 / KLK4 is secreted by transition- and maturation-stage ameloblasts. KLK4 aggressively degrades the retained organic matrix following the termination of enamel protein secretion. Two proteases are secreted into the enamel matrix of developing teeth. The early protease is enamelysin (MMP-2). The late protease is kallikrein 4 (KLK4). The principle functions of MMP-2 and KLK4 in dental enamel formation are to facilitate the orderly replacement of organic matrix with mineral, generating an enamel layer that is harder, less porous, and unstained by retained enamel proteins. Defects in Kallikrein-4 / KLK4 are the cause of amelogenesis imperfecta hypomaturation type 2A1 (AI2A1) which is an autosomal recessive defect of enamel formation. The disorder involves both primary and secondary dentitions.
Full Name
kallikrein-related peptidase 4
References
  • Nelson P.S., et al.,(1999), Molecular cloning and characterization of prostase, an androgen-regulated serine protease with prostate-restricted expression. Proc. Natl. Acad. Sci. U.S.A. 96:3114-3119.
  • Yousef G.M., et al., (1999), Prostase/KLK-L1 is a new member of the human kallikrein gene family, is expressed in prostate and breast tissues, and is hormonally regulated.Cancer Res. 59:4252-4256.
  • Stephenson S.A., et al.,(1999), Localization of a new prostate-specific antigen-related serine protease gene, KLK4, is evidence for an expanded human kallikrein gene family cluster on chromosome 19q13.3-13.4.J. Biol. Chem. 274:23210-23214.
  • Expression of Human Kallikreins 4, 8, 10, 11 and 13 in Pleomorphic Adenomas and Mucoepidermoid Carcinomas
    Author
    Hashemipour, MA;Fatah, FS;Ashraf, MJ;Tahmasebi, M;
    Year
    2016
    Journal
    Iranian Journal of Pathology
    Application
    control
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