The 4E2/MCP antibody reacts with mouse monocyte chemoattractant protein (MCP-1), formerly termed JE. The immunogen used to generate the 4E2/MCP hybridoma was mouse MCP-1. This antibody does not react with human MCP-1 and shows no reactivity with the closely related mouse β chemokines, TCA3 and MIP-1β.
ELISA Detection: The biotinylated 4E2/MCP antibody (Cat. No. 554444) is useful as a detection antibody for a sandwich ELISA for measuring mouse MCP-1 protein levels. Biotinylated 4E2/MCP antibody can be paired with a purified antibody such as the clone 2H5 as the capture antibody, with recombinant mouse MCP-1 (Cat. No. 554590) as the standard. Biotinylated 4E2/MCP antibody should be titrated 1.0 - 4.0 µg/ml to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of mouse MCP-1 ranging from ~4,000 to 30 pg/ml are recommended for inclusion in each ELISA plate. For maximal sensitivity, an overnight incubation (4°C) of samples/standards with the coated capture antibody is suggested. For specific methodology please visit the online protocols or the chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, G-CSF, GM-CSF, IFN-γ, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23 , SCF, TNF, LT-α, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF). Note 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring mouse MCP-1 in serum or plasma the BD OptEIA™ mouse MCP-1 ELISA Set (Cat. No. 555260) is specially formulated and recommended. Western Blot: The 4E2/MCP antibody has been found useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
