Human Cystatin C ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Cystatin C or cystatin 3 (formerly gamma trace, post-gamma-globulin or neuroendocrine basic polypeptide), a protein encoded by the CST3 gene, was originally described as a constituent of normal cerebrospinal fluid (CSF) and of urine from patients with renal failure.1 Cystatin 3 has a low molecular weight (approximately 13.3 kilodaltons), and it is removed from the bloodstream by glomerular filtration in the kidneys. In humans, all cells with a nucleus (cell core containing the DNA) produce cystatin C as a chain of 120 amino acids. It is found in virtually all tissues and bodily fluids. Cystatin C, which belongs to the type II cystatin gene family, is a potent inhibitor of lysosomal proteinases2 (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases3 (it prevents the breakdown of proteins outside the cell by a specific type of protein degrading enzymes) . Moreover, cystatin C is involved in network reorganization in the epileptic dentate gyrus.4

Expressed in submandibular and sublingual saliva but not in parotid saliva (at protein level) . Expressed in various body fluids, such as the cerebrospinal fluid and plasma. Expressed in highest levels in the epididymis, vas deferens, brain, thymus, and ovary and the lowest in the submandibular gland.

Apoptosis, Atherosclerosis, Cardiovascular, Caspases, Cell Biology, Cell Type Markers, Intracellular, Neural Stem Cells, Stem Cells, Tags & Cell Markers, Tumor Associated

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 687 2837 10374 647 2715 10112 Standard deviation 43.96 207.1 684.68 51.11 217.2 859.52 CV (%) 6.4% 7.3% 6.6% 7.9% 8% 8.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 687 2837 10374 647 2715 10112 Standard deviation 43.96 207.1 684.68 51.11 217.2 859.52 CV (%) 6.4% 7.3% 6.6% 7.9% 8% 8.5% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 687 2837 10374 647 2715 10112 Mean (pg/ml) 687 2837 10374 647 2715 10112 Standard deviation 43.96 207.1 684.68 51.11 217.2 859.52 Standard deviation 43.96 207.1 684.68 51.11 217.2 859.52 CV (%) 6.4% 7.3% 6.6% 7.9% 8% 8.5% CV (%) 6.4% 7.3% 6.6% 7.9% 8% 8.5%

As an inhibitor of cysteine proteinases, this protein is thought to serve an important physiological role as a local regulator of this enzyme activity.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0678-CA Rabbit anti- human CST3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0678-DA Biotinylated goat anti- human CST3 polyclonal antibody (Detection Antibody) 500 μl, 2.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0678-ST Lyophilized recombinant human CST3 standard 20 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0678-CA Rabbit anti- human CST3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0678-DA Biotinylated goat anti- human CST3 polyclonal antibody (Detection Antibody) 500 μl, 2.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0678-ST Lyophilized recombinant human CST3 standard 20 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0678-CA Rabbit anti- human CST3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0678-CA Rabbit anti- human CST3 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0678-DA Biotinylated goat anti- human CST3 polyclonal antibody (Detection Antibody) 500 μl, 2.3 μg/mL (recommended dilution 1:100) EZ0678-DA Biotinylated goat anti- human CST3 polyclonal antibody (Detection Antibody) 500 μl, 2.3 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0678-ST Lyophilized recombinant human CST3 standard 20 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0678-ST Lyophilized recombinant human CST3 standard 20 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.062 0.143 0.212 0.347 0.582 0.992 1.444 1.836 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.062 0.143 0.212 0.347 0.582 0.992 1.444 1.836 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 Concentration (pg/ml) 0 312 625 1250 2500 5000 10000 20000 O.D. 0.062 0.143 0.212 0.347 0.582 0.992 1.444 1.836 O.D. 0.062 0.143 0.212 0.347 0.582 0.992 1.444 1.836

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)