Cystatin C Rat (E. coli)

Description

Total 134 AA, Mw: 14.93 kDa (calculated). N-terminal His-tag, 14 extra AA (highlighted).

Amino Acid Sequence

Source

E. coli

Purity

>95%

SDS-PAGE gel

12% SDS-PAGE separation of Rat 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 0.03 M Acetate buffer, pH 4

Reconstitution

Add 0.1M Acetate buffer pH4 to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. For conversion into higher pH value, we recommend intensive dilution by relevant buffer to a concentration of 10μg/mL. In higher concentrations the solubility of this antigen is limited. 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 two weeks at 4°C.

Quality Control Test

BCA to determine quantity of the protein.

SDS PAGE to determine purity of the protein.

Applications

Western blotting

Note

This product is intended for research use only.

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.

References

• 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.
• 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.
• 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.
• 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.
• Price Ch.P., Finney H. Developments in the assessment of glomerular filtration rate. Clinica Chimica Acta 297, 55-66 (2000) .
• Ono S. et al. Increased cystatin C immunoreactivity in the skin in amyotrophic lateral sclerosis.
• 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) .
• 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) .
• 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.