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Recombinant fragment corresponding to Human MCP1.
QPDAINAPVT CCYNFTNRKI SVQRLASYRR ITSSKCPKEA VIFKTIVAKE ICADPKQKWV QDSMDHLDKQ TQTPKT
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 hMCAF/MCP-1 is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.|
|ELISA||Use a concentration of 0.5 µg/ml. To detect hMCAF/MCP-1 by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant hMCAF/MCP-1.|
|Neutralising||Use a concentration of 3 - 5 µg/ml. To yield one-half maximal inhibition [ND50] of the biological activity of hMCAF/MCP-1 (10.0 ng/ml), a concentration of 3.0 - 5.0 µg/ml of this antibody is required.|
|Sandwich ELISA||Use a concentration of 0.5 µg/ml.|
|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.
Image courtesy of Dr Laura Toma by Abreview.
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