Sandwich ELISA analysis of Human IL-17A monoclonal antibody was performed using the Thermo Scientific ELISA Kit (Product # EHIL17A) by coating a blank 96-well microtiter plate with 100µl per well of Human IL-17A moncolonal antibody (Product # MA1-068) in duplicate at 1, 3, 4, 5, 7, and 9µg/ml in DPBS (Product # 28374) and incubating for 12-18 hours at 4C. The plate was aspirated and blocked with 300µl per well of 4% BSA and 5% sucrose in DPBS for 1 hour at room temperature. Human IL-17A recombinant protein at 50µl per well was added in duplicate at 2000, 1000, 500, 250, 125, 62.5, 31.25, and 0 pg/ml for 2 hours at room temperature along with 50µl of Human IL-17A biotinylated monoclonal antibody (Product # MA1-069B) in all applicable wells at 0.1µg/ml for 2 hours at room temperature. The plate was washed with ELISA Wash Buffer (Product # N503) and incubated with 100µl per well of Streptavidin-HRP (Product # N504) in all test wells at 1:40,000 dilution for 30 minutes at room temperature and then washed and incubated with 100µl per well of TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid (Product # N600). Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant Human IL17A.|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1 - 3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The MA1068 anti-IL17a antibody (clone 3-220.127.116.11) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody MA1069B (biotinylated conjugate of clone 3-18.104.22.168). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125-0.25 µg/ml for detection.
The protein encoded by IL-17 gene is a proinflammatory cytokine produced by activated T cells. This cytokine regulates the activities of NF-kappaB and mitogen-activated protein kinases.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.