Human Kallikrein 8 HEK293 Overexpression Lysate: 製品の情報
This Human Kallikrein 8 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Kallikrein 8 protein (Cat: 11820-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human KLK8 isoform 1 (O60259-1) (Met 1-Gly 260) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant human KLK8 consists of 243 amino acids and predictes a molecular mass of 26.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhKLK8 is approximately 36 kDa due to glycosylation.
Human Kallikrein 8 HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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 Kallikrein 8 HEK293 Overexpression Lysate: 別名
Human HNP Overexpression Lysate; Human NP Overexpression Lysate; Human NRPN Overexpression Lysate; Human PRSS19 Overexpression Lysate; Human TADG14 Overexpression Lysate
Kallikrein 8 背景情報
Kallikrein-8, also known as Neuropsin, Serine protease 19, Serine protease TADG-14, Tumor-associated differentially expressed gene 14 protein and KLK8, is a secreted protein which belongs to thepeptidase S1 family and Kallikrein subfamily. It is a serine protease which is capable of degrading a number of proteins such as casein, fibrinogen, kininogen, fibronectin and collagen type IV. Kallikrein-8 / KLK8 plays a role in the formation and maturation of orphan and small synaptic boutons in the Schaffer-collateral pathway. It regulates Schaffer-collateral long-term potentiation in the hippocampus and is required for memory acquisition and synaptic plasticity. It is involved in skin desquamation and keratinocyte proliferation and plays a role in the secondary phase of pathogenesis following spinal cord injury. It also cleaves L1CAM in response to increased neural activity. It induces neurite outgrowth and fasciculation of cultured hippocampal neurons. Kallikrein-8 / KLK8 is expressed at high levels in serum, ascites fluid and tumor cytosol of advanced stage ovarian cancer patients and may serve as a marker of ovarian cancer. Kallikrein-8 / KLK8 may have potential clinical value for disease diagnosis or prognosis and it may also be a useful therapeutic target.
Yoshida S., et al.,(1998), Sequence analysis and expression of human neuropsin cDNA and gene. Gene 213:9-16.
Underwood L.J., et al., (1999), Cloning of tumor-associated differentially expressed gene-14, a novel serine protease overexpressed by ovarian carcinoma.Cancer Res. 59:4435-4439.
Mitsui S., et al.,(1999), A novel form of human neuropsin, a brain-related serine protease, is generated by alternative splicing and is expressed preferentially in human adult brain.Eur. J. Biochem. 260:627-634.
Expression of Human Kallikreins 4, 8, 10, 11 and 13 in Pleomorphic Adenomas and Mucoepidermoid Carcinomas
Hashemipour, MA;Fatah, FS;Ashraf, MJ;Tahmasebi, M;