Canine Cystatin C ELISA Assay Kit
The Canine Cystatin C ELISA Assay Kit is For Research Use Only
Size: 1×96 wells
Dynamic Range: 0.07 ng/mL – 4 ng/mL
Incubation Time: 1 hour 50 minutes
Sample Type: Plasma, Serum, Urine
Sample Size: 100 μL
Alternative Names: Cystatin C
Assay Principle
The principle of the double antibody sandwich ELISA is represented in Figure 1. In this assay the Cystatin C (CYS) present in samples reacts with the anti-CYS antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, the Detection Antibody, biotin conjugated anti- CYS is added and complexes are formed. Following a wash step, the horseradish peroxidase (HRP) conjugated Streptavidin is added and complexes are formed. After another washing step, the complexes are assayed by the addition of a chromogenic substrate, 3,3’,5,5’-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of CYS in the sample tested; thus, the absorbance, at 450 nm, is a measure of the concentration of CYS in the test sample. The quantity of CYS in the test sample can be interpolated from the standard curve constructed from the standards and corrected for sample dilution.
Specimen Collection
- Serum samples – Blood should be collected by venipuncture. The serum should be separated from the cells after clot formation by centrifugation. Remove serum and assay immediately or aliquot and store samples at –80oC (preferably) or -20oC. Avoid repeated freeze-thaw cycles.
- Plasma samples – Blood should be collected into a container with an anticoagulant and then centrifuged. Assay immediately or aliquot and store samples at –80oC (preferably) or -20oC. Avoid repeated freeze-thaw cycles.
- Urine samples – Collect mid-stream using sterile or clean urine collector. Centrifuge to remove cell debris. Assay immediately or aliquot and store samples at –80oC (preferably) or -20oC. Avoid repeated freeze-thaw cycles.
- Known interfering substances – Azide and thimerosal at concentrations higher than 0.1% inhibits the enzyme reaction.
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