Preparation and Storage
推奨アッセイ手順
ELISA Capture: The purified BVD2-23B6 antibody (Cat. No. 554502) is useful as a capture antibody in a sandwich ELISA for measuring human GM-CSF protein levels. The purified BVD2-23B6 antibody can be paired with the biotinylated BVD2-21C11 antibody (Cat. No. 554505) as the detection antibody, with recombinant human GM-CSF (Cat. No. 550068) as the standard. Purified BVD2-23B6 antibody should be titrated 2 - 6 µ/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of GM-CSF standard ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate.
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, MCP-1, TCA-3, TNF-α; 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, 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).
This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum samples. For testing human GM-CSF in complex biological fluids such as serum or plasma, our Human GM-CSF BD OptEIA™ ELISA Set (Cat. No. 555126) is recommended.
Neutralization: The NA/LE format of the BVD2-23B6 antibody (Cat. No. 554501) is useful for neutralization of human GM-CSF bioactivity. A suitable NA/LE. rat IgG2a isotype control is the R35-95 antibody, (Cat. No. 554687).
IP/WB: The BVD2-23B6 antibody (e.g., Cat. No. 554502) has been reported to be useful for immunoprecipitation studies. For Western blotting, a concentration of 1 µ/ml has been found to enable visualization of ≤ 100 ng/lane of recombinant human GM-CSF (e.g., Cat. No. 550068), under reducing conditions.
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 BVD2-23B6 antibody reacts with human granulocyte/macrophage - colony stimulating factor (GM-CSF). The immunogen used to generate the BVD2-23B6 hybridoma was E. coli-expressed recombinant human GM-CSF. This is a neutralizing antibody.
This antibody is routinely tested by ELISA analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
Development References (4)
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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, Immunoprecipitation, Neutralization). View Reference
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Abrams JS, Silver JE, Van Dyke RE, Gleich GI. Eosinophil-active cytokines in human disease: development and use of monoclonal antibodies to IL-3, IL-5 and GMCSF. In: Gleich GJ and Kay AB, ed. Eosinophils in Allergy and Inflammation. New York: Dekker; 1994:133-157.
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Bacchetta R, de Waal Malefijt R, Yssel H. Host-reactive CD4+ and CD8+ T cell clones isolated from a human chimera produce IL-5, IL-2, IFN-gamma and granulocyte/macrophage-colony-stimulating factor but not IL-4. J Immunol. 1990; 144(3):902-908. (Clone-specific: ELISA, Neutralization). View Reference
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Kita H, Ohnishi T, Okubo Y, Weiler D, Abrams JS, Gleich GJ. Granulocyte/macrophage colony-stimulating factor and interleukin 3 release from human peripheral blood eosinophils and neutrophils. J Exp Med. 1991; 174(3):745-748. (Clone-specific: ELISA, 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.
