MCP-4 Antibody (M810) in ELISA-2<br />Sandwich ELISA analysis of an anti-human MCP-4 monoclonal antibody (Product # M809) was performed by loading 100ul per well of antibody (Product # M809) at a concentration of 2ug/ml overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100ul of recombinant human MCP-4/CCL13 was added to wells in duplicate at 2000, 500, 125, 31.25, 7.813, and 0pg/ml concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100ul per well of a biotinylated MCP-4 monoclonal antibody (Product # M810, biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435)) at a concentration of 0.13ug/ml for 1 hour at room temperature, followed by 100ul per well of Streptavidin-HRP (Product # N504) at a dilution of 1:16,000 for 30 minutes at room temperature. Detection was performed by adding 100ul of Ultra TMB substrate (Product # 34028) per well and incubating for 30 minutes at room temperature in the dark. The plate was then stopped with 100ul per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.<br />
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant CCL13 protein|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||2 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 M810 MCP-4 antibody (clone 8C12) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M809 (biotinylated conjugate of clone 3G4). The M810 MCP-4 antibody (biotinylated conjugate of clone 8C12) has also been used successfully in a sandwich ELISA as a detection antibody when paired with M809 (clone 3G4) as the coating antibody.
Cytokines are a family of secreted proteins involved in immunoregulatory and inflammatory processes. The gene encoding MCP-4/CCL13 is one of several Cys-Cys (CC) cytokine genes clustered on the q-arm of chromosome 17. The CC cytokines are proteins characterized by two adjacent cysteines. The cytokine encoded by this gene displays chemotactic activity for monocytes, lymphocytes, basophils and eosinophils, but not neutrophils. MCP-4/CCL13 signals through CCR2B and CCR3 receptors, and plays a role in the accumulation of leukocytes at both sides of allergic and non-allergic inflammation. It be also involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis, and may also play a role in monocyte attraction in tissues chronically exposed to exogenous pathogens.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.