Human Kallikrein 13 HEK293 Overexpression Lysate 产品信息
Product Description
This Human Kallikrein 13 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Kallikrein 13 protein (Cat: 10199-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the pro-form of human KLK13 (NP_056411.1) (Met 1-Ile 262) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human KLK13 consists of 256 amino acids and has a predicted molecular mass of 28.4 kDa. As a result of glycosylation, the apparent molecular mass of rh KLK13 is approximately 33-37 kDa in SDS-PAGE under reducing conditions.
Human Kallikrein 13 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 Kallikrein 13 HEK293 Overexpression Lysate Alternative Names
Human KLK-L4 Overexpression Lysate;Human KLKL4 Overexpression Lysate
Kallikrein 13 Background Information
Tissue kallikrein 13 (hK13), also known as KLK-L4 (kallikrein-like gene 4), is a member of the human tissue kallikrein family of serine proteases having diverse physiological functions in many tissues. The KLK13 gene resides on chromosome 19q13.3-4 along with other 14 members in a gene cluster and shares a high degree of homology. KLK13 is a trypsin-like, secreted serine protease expressed specifically in the testicular tissue including prostate, salivary gland, breast, and testis. Growing evidence suggests that many kallikreins are implicated in carcinogenesis and may play a role in metastasis. KLK13 may be involved in the pathogenesis and/or progression of breast and ovary cancers, and is regarded as a novel cancer biomarker. In addition, KLK13 interacts and forms complexes with several serum protease inhibitors, such as alpha2-macroglobulin, and its expression is regulated by steroid hormones.
Full Name
kallikrein-related peptidase 13
References
Yousef G.M., et al.,(2000), Identification and characterization of KLK-L4, a new kallikrein-like gene that appears to be down-regulated in breast cancer tissues. J. Biol. Chem. 275:11891-11898. Komatsu N., et al., (2003), Expression and localization of tissue kallikrein mRNAs in human epidermis and appendages.J. Invest. Dermatol. 121:542-549. Andres A.M., et al.,(2007), Understanding the accuracy of statistical haplotype inference with sequence data of known phase.Genet. Epidemiol. 31:659-671.