Cystatin C Canine E. coli

Introduction to the Molecule

Cystatin C is a non-glycosilated basic single-chain protein consisting of 120 amino acids with a molecular weight of 13.36 kDa and is characterized by two disulfide bonds in the carboxy-terminal region. It belongs to the cystatins superfamilly which inactivates lysosomal cysteine proteinases, e.g. cathepsin B, H,.K, L and S. Imbalance between Cystatin C and cysteine proteinases is associated with inflammation, renal failure, cancer, Alzheimer's di­sease, multiple sclerosis and hereditary Cystatin C amyloid angiopathy. Its increased level has been found in patients with autoimune diseases, with colorectal tumors and in patients on dyalisis. Serum Cystatin C seems to be better marker of glomerular filtration rate than creatinine. On the other hand, low concentration of Cystatin C presents a risk factor for secondary cardiovascular events.

Research topic

Renal disease

Description

Total 133 AA. MW: 14,85 kDa (calculated). N-Terminal His-tag, 8 extra AA (highlighted).

Amino Acid Sequence

Source

E. coli

SDS-PAGE gel

14% SDS-PAGE separation of Canine Cystatin C 1. M.W. marker – 14, 21, 31, 45, 66, 97 kDa 2. reduced and heated sample, 5μg / lane 3. non-reduced and non-heated sample, 5μg / lane

Formulation

Filtered (0,4 μm) and lyophilized in 0.5 mg/mL in 20mM TRIS, 50mM NaCl, pH 7.5

Reconstitution

Add deionized water to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. Product is not sterile! Please filter the product by an appropriate sterile filter before using it in the cell culture.

Storage, Stability/Shelf Life

Store lyophilized protein at –20°C. Lyophilized protein remains stable until the expiry date when stored at –20°C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80°C for long term storage. Reconstituted protein can be stored at 4°C for a limited period of time; it does not show any change after one week at 4°C.

Quality Control Test

BCA to determine quantity of the protein.

SDS PAGE to determine purity of the protein.

Applications

Western blotting

References

• Bokenkamp A, Ciarimboli G, Dieterich C. Cystatin C in a rat model of end-stage renal failure. Ren Fail. 2001 May-Jul;23(3-4):431-8.
• Lindahl P., Abrahamson M., Bjork L.:. Interaction of recombinant human cystatin C with the cysteine proteinase papain and actinidin. Biochem. J. 281, 49-55 (1996) .
• Nagai A, Ryu JK, Kobayash S, Kim SU. Cystatin C induces neuronal cell death in vivo. Ann N Y Acad Sci. 2002 Nov;977:315-21.
• Nicklin M.J.H. and Barrett A.J. Inhibition of cysteine proteinases and dipeptidyl peptidase 1 by eggwhite cystatin. Biochem J. 223, 245-253 (1984) .
• Ono S. et al. Increased cystatin C immunoreactivity in the skin in amyotrophic lateral sclerosis.
• Page M.K. et al.:. Clinical value of cystatin C determination. Clinica Chim. Acta 297, 67-72 (2000) .
• Pirttila TJ, Pitkanen A. Cystatin C expression is increased in the hippocampus following photothrombotic stroke in rat. Neurosci Lett. 2006 Mar 6;395(2):108-13. Epub 2005 Nov 23.
• Price Ch.P., Finney H. Developments in the assessment of glomerular filtration rate. Clinica Chimica Acta 297, 55-66 (2000) .
• Roald AB, Aukland K, Tenstad O. Tubular absorption of filtered cystatin-C in the rat kidney. Exp Physiol. 2004 Nov;89(6):701-7. Epub 2004 Aug 24.
• Xu L, Sheng J, Tang Z, Wu X, Yu Y, Guo H, Shen Y, Zhou C, Paraoan L, Zhou J. Cystatin C prevents degeneration of rat nigral dopaminergic neurons: in vitro and in vivo studies. Neurobiol Dis. 2005 Feb;18(1):152-65.

By research topic:

• Renal disease

By molecule:

• Cystatin C