| Cystatin B(CSTB), also called STFB, is a small protein that is a member of the superfamily of cysteine protease inhibitors. It has been isolated from human spleen and liver and its amino acid sequence has been fully determined. The cystatin B gene is located on 21q22.3. It is widely distributed and is localized mostly intracellularly, but has been found extracellularly. The protein is able to form a dimer stabilized by noncovalent forces, inhibiting papain and cathepsins l, h and b. Its role is thought to be as a protector against the proteinases leaking from lysosomes. A cystatin B multiprotein complex might have a specific cerebellar function, and that the loss of this function might contribute to the etiopathogenesis of EPM1. Upon differentiation to myotubes, CSTB becomes excluded from the nucleus and lysosomes, suggesting that the subcellular distribution of CSTB is dependent on the differentiation status of the cell. Sample Type: Cell culture supernates, serum, plasma (heparin, EDTA), saliva, urine and human milk Intended Use: For quantitative detection of human Cystatin B in cell culture supernates, serum, plasma (heparin, EDTA), saliva, urine and human milk. Sensitivity: <10pg/ml Range: 62.5-4000pg/ml Specificity: Natural and recombinant human Cystatin B Test Principle: This human Cystatin B BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Cystatin B has been precoated onto 96-well plates. Standards(E.coli) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Cystatin B is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Cystatin B amount of sample captured in plate. Kit Components: 96-well plate precoated with human Cystatin B antibody, 1 Lyophilized recombinant human Cystatin B standard, 10ng/tube*2 Biotinylated human Cystatin B antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
