Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | Human FB Proteolysis Assay (ELISA C3a) |
| Description: | Human CVF-Bb complex (3 nM concentration) was incubated with compound at various concentrations for 1 h at room temperature in PBS at pH 7.4, containing 10 mM MgCl2 and 0.05% (w/v) CHAPS. The enzyme reaction was started by addition of C3 diluted in the assay buffer to a final concentration of 1 μM. After 1 h incubation time at room temperature, the enzyme reaction was stopped by addition of an excess of various enzyme inhibitors (Roche, cOmplete Inhibitor Tablets). Generation of C3a as the enzymatic cleavage product was quantified by an enzyme-linked immunosorbent assay (ELISA). Aliquots of reaction samples were pipetted into 384-well high-capacity protein-binding plates (NUNC MaxiSorp) pre-filled with 97 μL/well of coating buffer. After an overnight incubation at 4 °C, assay plates were washed with PBS-Tween 20. Remaining free binding capacity was saturated by the addition of Starting Block T20 for 5 min at room temperature, and assay plates were then washed with PBS-Tween 20. Anti-C3a neoepitope antibody was added to each well, followed by incubation for 60 min at room temperature. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |
|
Home |
| |
Search |
| |
Deposit |
| |
SiteMap |
| |
About us |
| |
Email us |
| |
Info |
|
©2000 BindingDB. All rights reserved. |
|||||||||||||