The Aviva Rat IP-10ELISA Kit contains the components necessary for quantitative determination of natural or recombinant rIP-10concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator.The capture antibodies coated to the bottom of each well are specific for a particular epitope on theRatIP-10cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetricreaction to ensue upon substrate addition. When the substrate TMB (3, 3’, 5, 5’-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
ELISA Kit Range
16-1000 pg/mL
ELISA Kit Component
Components
Amount
Capture Antibody Coated Microplate
96 Wells (12 x 8)
Ready-to-Use Streptavidin-HRP
12 mL
Ready-to-Use Substrate
12 mL
Stop Solution
12 mL
Wash Buffer (15x)
50 mL
Protein Standard Diluent
12 mL
Sample Diluent
12 mL
Detection Antibody Diluent
12 mL
Biotin-Labeled Detection Antibody
Lyophilized
Protein Standard
Lyophilized (100 ng)
Reconstitution and Storage
2°C to 8°C
Sample Type
Cell lysates, sera and plasma
Sensitivity
16 pg/mL
Specificity
The Rat IP-10 ELISA is capable of recognizing both recombinant and naturally produced Rat IP-10 proteins. The antigens listed below were tested at 50 ng/ml and exhibited less than 2% cross reactivity. • Murine: IP-10 The antigens listed below were tested at 50 ng/ml and did not exhibit significant cross reactivity or interference. • Human: ENA-78, GCP-2, IL-8 (72 aa), IL-8 (77 aa), IP-10, I-TAC, MIG, NAP-2 • Murine: BCA-1/BLC, I-TAC, MIG, SDF-1α, SDF-1β • Rat: KC