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Active Cystatin C (Cys-C)

CST3; Cystatin 3; Gamma Trace; Post-Gamma-Globulin; Neuroendocrine Basic Polypeptide; Amyloid Angiopathy And Cerebral Hemorrhage

ACTIVITY TEST

Cystatin C is a member of family 2 of the Cystatin superfamily. It is involved in processes such as tumor invasion and metastasis, inflammation and some neurological diseases. It inhibits many cysteine proteases such as papain and cathepsins B, H, K, L and S. It is ubiquitous in human tissues and body fluids. A point mutation in the gene coding for the 120 amino acid mature Cystatin C causes a hereditary form of amyloid angiopathy in which the protein variant (Leu68 to Gln) is deposited in the cerebral arteries, leading to fatal cerebral hemorrhage. Cystatin C may have additional clinical applications. For example, it is a good marker for glomerular filtration rate. The activity of recombinant rat Cystatin C was measured by its ability to inhibit papain cleavage of a fluorogenic peptide substrate Z-FR-AMC in the assay buffer 50 mM Tris, pH 7.0. Papain was diluted to 500 ug/ml in activation buffer 50 mM Tris, 5 mM DTT, pH 7.0 and incubated at room temperature for 15 minutes. The activated papain was diluted to 100 ug/ml in the assay buffer and 20 ul different concentrations of recombinant rat Cystatin C (MW: 14.93 KD) was incubated with 20 ul 100 ug/ml papain at 37 ℃ for 10 minutes. Loading 50 µL of the incubated mixtures which were diluted five-fold in assay buffer into empty wells of a plate, and start the reaction by adding 50 µL of 200 µM substrate. Include a substrate blank containing 50 µL of assay buffer and 50 µL of 200 µM substrate. Then read at excitiation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes. The result was shown in Figure 1 and it was obvious that recombinant rat Cystatin C significantly decreased papain activity. The inhibition IC50 was <80 nM.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
BioMed Research International The Renal Effects of Vanadate Exposure: Potential Biomarkers and Oxidative Stress as a Mechanism of Functional Renal Disorders—Preliminary Studies Hindawi: 740105
Critical Care Endothelial dysfunction and renal fibrosis in endotoxemia-induced oliguric kidney injury: possible role of LPS-binding protein Pubmed:25261195
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Journal of Agricultural and Food Chemistry Protective Effects of DHA-PC against Vancomycin-Induced Nephrotoxicity through the Inhibition of Oxidative Stress and Apoptosis in BALB/c Mice pubmed:29254330
THERAPEUTIC ADVANCES IN RESPIRATORY DISEASE Association of cathepsin B and cystatin C with an age-related pulmonary subclinical state in a healthy Chinese population Pubmed: 32401159
RSC Adv SERS based Y-shaped aptasensor for early diagnosis of acute kidney injury Pubmed:35733690
Food Funct A Compared Study of Gentle Ketogenic Diet Containing Medium-Chain Triglycerides or Long-Chain Triglycerides on Chronic Sleep Deprivation-Induced Cognitive … Pubmed:35141738
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPA896Ra01 Recombinant Cystatin C (Cys-C) Positive Control; Immunogen; SDS-PAGE; WB.
APA896Ra61 Active Cystatin C (Cys-C) Cell culture; Activity Assays.
EPA896Ra61 Eukaryotic Cystatin C (Cys-C) Positive Control; Immunogen; SDS-PAGE; WB.
PAA896Ra01 Polyclonal Antibody to Cystatin C (Cys-C) WB; IHC; ICC; IP.
LAA896Ra71 Biotin-Linked Polyclonal Antibody to Cystatin C (Cys-C) WB; IHC; ICC.
MAA896Ra21 Monoclonal Antibody to Cystatin C (Cys-C) WB; IHC; ICC; IP.
SEA896Ra ELISA Kit for Cystatin C (Cys-C) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA896Ra Multiplex Assay Kit for Cystatin C (Cys-C) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.