ID | Components | Size |
EP135-C01 | High-bind Plate | 1 plate |
EP135-C02 | Human CD30L | 15 μg |
EP135-C03 | Biotinylated Human CD30 | 10 μg |
EP135-C04 | Anti-CD30 Neutralizing Antibody | 20 μg |
EP135-C05 | Streptavidin-HRP | 10 μg |
EP135-C06 | Coating Buffer | 12 mL |
EP135-C07 | 10xWashing Buffer | 50 mL |
EP135-C08 | Blocking Buffer | 50 mL |
EP135-C09 | Substrate Solution | 12 mL |
EP135-C10 | Stop Solution | 7 mL |
背景(Background)
CD30 belongs to the tumor necrosis factor receptor (TNFR) superfamily, and the physiological functions of CD30 are in addition to the transmission of synergistic stimulation signals. CD30 ligand (CD30L) belongs to the tumor necrosis factor (TNF) superfamily, and CD30L is present in activated T cells, quiescent B cells, granulocytes, thymic medullary cells, and various leukemia cells. After crosslinking CD30 and CD30L, diverse physiological function signaling is led by TNFR correlation factors (TRAF), including TRAF1, 2, 3, 5. CD30 was originally found in Hodgkin's lymphoma and Reed Sternberg (RS) cells, and is now known to be expressed on the surface of some normal lymphocytes, virally infected lymphocytes, lymphocytes of lymphogenetic origin, and activated T cell subsets that produce Th2 cytokines.
应用说明(Application)
This kit is developed for screening for inhibitors of human CD30 binding to human CD30L.
It is for research use only.
重构方法(Reconstitution)
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(Storage)
Unopened kit should be stored at 2°C -8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理(Assay Principles)
This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new CD30 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human CD30 to immobilized human CD30 Ligand in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated CD30 protein, a human CD30 Ligand protein, an anti-CD30 neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human CD30 Ligand.
b) Add your molecule of interest to the tests.
c) Add human CD30-Biotin to bind the coated CD30 Ligand.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to CD30 Ligand: CD30 binding will be determined by comparing OD readings among different experimental groups.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
INHIBITION OF CD30[Biotinylated]: CD30L BINDING BY ANTI-CD30 NEUTRALIZING ANTIBODY
Serial dilutions of Anti-CD30 Neutralizing antibody (Catalog # EP135-C04) (1:1 serial dilution, from 5 μg/mL to 0.019μg/mL) was added into Biotinylated CD30: CD30L binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).