Human Cystatin F / CST7 HEK293 Overexpression Lysate

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Human Cystatin F / CST7 HEK293 Overexpression Lysate: Product Information

Product Description
This Human Cystatin F / CST7 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Cystatin F / CST7 protein (Cat: 10167-H02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human Cystatin F precursor (NP_003641.3) (Met 1-His 145) was expressed with the fused Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human Cystatin-F/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 364 amino acids and predicts a molecular mass of 41.3 kDa. As a result of glycosylation, the rh Cystatin-F/Fc monomer migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.

Human Cystatin F / CST7 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 Cystatin F / CST7 HEK293 Overexpression Lysate: Alternative Names

Human CMAP Overexpression Lysate

Cystatin F / CST7 Background Information

The cystatin superfamily members are important natural cysteine protease inhibitors present in a wide variety of organisms and are divided into three classes. Cystatin F, also known as leukocystatin and CMAP (Cystatin-like Metastasis-Associated Protein), is a type 2 cystatin and its expression is limited to hematopoietic cells, with the highest expression levels being observed in monocytes, dendritic cells, and certain types of T-cells. Furthermore, cystatin F mRNA becomes up-regulated during dendritic cell maturation, and thus suggests a specific role of cystatin F in immune regulation. Cystatin F is produced as a dimer, an inactive cathepsin inhibitor which is activated by chemical reduction. In addition, Cystatin F and its homologues have been observed expressing in various human cancer cell lines established from malignant tumors, and thus indicates a new diagnosis and prevention approach of certain human carcinomas metastasis.
Full Name
cystatin F (leukocystatin)
References
  • Halfon S., et al.,(1998), Leukocystatin, a new class II cystatin expressed selectively by hematopoietic cells. J. Biol. Chem. 273:16400-16408.
  • Ni J., et al., (1998), Cystatin F is a glycosylated human low molecular weight cysteine proteinase inhibitor.J. Biol. Chem. 273:24797-24804.
  • Morita M., et al.,(2000), Genomic construct and mapping of the gene for CMAP (leukocystatin/cystatin F, CST7) and identification of a proximal novel gene, BSCv (C20orf3).Genomics 67:87-91.
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