Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified MP1-22E9 antibody (Cat. No. 554404) is useful as a capture antibody for a sandwich ELISA for measuring mouse GM-CSF protein levels. Purified MP1-22E9 antibody can be paired with the biotinylated MP1-31G6 (Cat. No. 554407) antibody as the detecting antibody, with recombinant mouse GM-CSF (Cat. No. 554586) as the standard. Purified MP1-22E9 antibody should be titrated 1-4 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse GM-CSF ranging from ~2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1ß, 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-1ß, 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 GM-CSF in serum or plasma our mouse GM-CSF BD OptEIA™ set (Cat. No. 555167) is specially formulated and recommended.
Western Blot: The MP1-22E9 antibody (Cat. 554404) has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Neutralization: The NA/LE™ MP1-22E9 antibody (Cat. No. 554403) is useful for neutralization of mouse GM-CSF bioactivity.
Immunofluorescent Staining and Flow Cytometric Analysis: The MP1-22E9 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify and enumerate GM-CSF producing cells within mixed cell populations. The PE-conjugated MP1-22E9 antibody (Cat. No. 554406) is especially suitable for these experiments.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
The MP1-22E9 monoclonal antibody specifically binds to mouse Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF). The immunogen used to generate the MP1-22E9 hybridoma was yeast-expressed recombinant mouse GM-CSF. This is a neutralizing antibody.
Development References (5)
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Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). View Reference
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Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA, Neutralization). View Reference
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Nozaki S, Abrams JS, Pearce MK, Sauder DN. Augmentation of granulocyte/macrophage colony-stimulating factor expression by ultraviolet irradiation is mediated by interleukin 1 in Pam 212 keratinocytes. J Invest Dermatol. 1991 July; 97(1):10-14. (Clone-specific: ELISA, Neutralization). View Reference
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Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific: ELISA). View Reference
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Suda T, O'Garra A, MacNeil I, Fischer M, Bond MW, Zlotnik A. Identification of a novel thymocyte growth-promoting factor derived from B cell lymphomas. Cell Immunol. 1990; 129(1):228-240. (Clone-specific: Neutralization). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.