Biolegend, 756224, Cell-Vive™ GMP Recombinant Human Cystatin C (carrier-free), 50μg

A member of the the cystatin sub-family, Cystatin C was initially identified in human urine and named gamma-trace. The cystatin family contains three subfamilies: Type 1 cystatins, also known as stefins, which are predominantly present in the cytosol and the nuclei; Type 2 cystatins are mainly secreted proteins; Type 3 cystatins are multi-domain proteins of high molecular weight (60-120 kD) and are also known as kininogens. Cystatin C inhibits cysteine proteases from the papain family and legumain. It forms tight non-covalent associations with its target enzymes. Cystatin C antagonizes TGF-β signaling in normal and cancer cells in vitro . In this process, Cystatin C binds to the TGF-β type II receptor and inhibits the binding of TGF-β. Cystatin C influences the progression of diseases such as, cancer metastasis, atherosclerosis, aortic aneurisms, emphysema, and arthritis. Also, it is associated with neurodegenerative disease and cerebral angiopathy. Human Cystatin C has the tendency to dimerize in order to produce very tight two-fold symmetric dimers. This amyloidogenic property suggests a mechanism for its aggregation in the brain arteries of elderly people with amyloid angiopathy. The L68Q mutant of human Cystatin C is responsible for conformational disease, which causes massive amyloidosis, cerebral hemorrhage, and death in young adults. BioLegend Cell-Vive™ GMP Recombinant proteins are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Ph. Eur. Chapter 5.2.12 in a dedicated GMP facility compliant with ISO 13485:2016. Specifications and processes include:
50μg
Source: Human Cystatin C, amino acids Ser27-Ala146 (Accession# P01034) was expressed in 293E cells. The carboxy-terminal contains a TG-10His-GGQtag.
Molecular Mass: The 135 amino acid recombinant protein has a predicted molecular mass of 15.1 kD. The non-reduced and DTT-reduced protein migrates at approximately 17 kD by SDS-PAGE. The predicted N-terminal amino acid is Serine.
N-terminalSequence Analysis: Ser-Ser-Pro-Gly-Lys-Pro-Pro-Arg-Leu-Val
Purity: ≥ 95%, as determined by Coomassie stained SDS-PAGE
Formulation: Protein was lyophilized from 0.1 μm filtered solution containing 20mM HEPES, 0.1M NaCl, pH7.2
Endotoxin Level: Less than or equal to 0.1 EU per μg of protein as determined by LAL method.
Residual Host Cell Protein Content: ≤ 0.50 ng/μg by ELISA
Concentration: 50 μg and 500 μg sizes are lyophilized
Storage & Handling: Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at - 20°C or colder until the expiration date. Reconstitute lyophilized protein in sterile water. Before reconstitution, make sure sterile water and product are at room temperature. Quickly spin the vial or gently tap down on the vial to make sure the lyophilized product is at the bottom of the vial before opening. Use aseptic techniques to add the required volume of reconstitution buffer (sterile water) to the vial, to obtain the recommended stock concentration 250 μg/mL. Close the vial and leave at ambient temperature for 15-20 minutes. Then gently invert the vial several times or until all of the lyophilized product dissolves. Leave the vial at room temperature for another 15 minutes. If small particulates are still observed after 15 minutes, incubate at room temperature for an additional 30 minutes and leave the vial at 2°C - 8°C overnight. Next day, invert the vial several times or gently pipette the solution up and down before use. If needed, transfer the reconstituted stock solution to a sterile container for additional dilution to no less than 100 μg/mL. Small working aliquots in polypropylene tubes can be made after reconstitution and store the vials at -20°C or lower. Avoid freeze/ thaw cycles. Carrier protein such as 0.2 - 1% endotoxin-free BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to two weeks or stored at -20°C or colder for up to 3 months.
Activity: IC50 < 6.0 nM, as determined by inhibiting human Papain (0.05 μg/mL) activity when 100 µM of Z-FR-AMC is used as a papain peptide substrate.
Application: Bioassay
Application Notes: Our lyophilized proteins are validated in-house to maintain activity after shipping at ambient temperature and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
Disclaimer: BioLegend Cell-Vive™ GMP Recombinant proteins are for research use only. Suitable for ex vivo cell processing. Not for injection or diagnostic or therapeutic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.
Structure: Monomer
Function: Inhibitor of cysteine proteases (capthepsins B, H, K, L, and S). Transcription factor IFN regulatory factor 8 (IRF-8) induces cystatin C in mouse primary dendritic cells. IL-10 inhibits IRF-8 and consequently Cystatin C.
Interaction: Normal and cancer cells.
Ligand/Receptor: Papain-like cysteine proteases, cathepsins B, H, K, L, and S, and legumain.
Bioactivity: Cystatin C inhibits the activity of papain.
Cell Sources: Considered ubiquitously expressed, biological fluids, dendritic cells, and macrophages.
Antigen References: Grubb A, et al. 1982. Proc. Nat. Acad. Sci. 79:3024. Janowski R, et al. 2001. Nat. Struct. Biol. 8:316. Sokol JP, Schiemann WP. 2004. Mol. Cancer Res. 2:183. Bird PI, et al. 2009. Nat Rev Immunol. 9:871. Kaur G, Levy E. 2012. Front Mol. Neurosci. 5:79. Xu Y, et al. 2014. J Biol Chem. 289:9730. Xu Y, et al. 2015. Immunol. Cell Biol. 5:442.
Gene ID: 1471
UniProt: View information about Cystatin C on UniProt.org
Other Names: Cystatin-3, Gamma-trace, Neuroendocrine basic polypeptide, Post-gamma-globulin, gamma-CSF, post-gamma protein