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Our Abpromise guarantee covers the use of ab9669 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.1 - 0.2 µg/ml.
To detect hMCAF/MCP-1 by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant MCP-1 is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.
|Sandwich ELISA||Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal to MCP1 (ab9858).
To detect MCP-1 by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of ab9669 is required. This antigen affinity purified antibody, in conjunction with ab9858 as a capture antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant MCP-1.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. PubMed: 20369226|
HL-60 cells were incubated at 37°C for 24hrs with vehicle control (0 µM) and varied concentrations of 2-Arachidonylglycerol (ab120098). Increased expression of MCP1 in HL-60 cells correlates with an increase in 2-Arachidonylglycerol concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab9669 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.