ELISA Detection: The biotinylated BVD2-21C11 antibody (Cat. No. 554505) is useful as a detection antibody in a sandwich ELISA for measuring human GM-CSF protein levels. Biotinylated BVD2-21C11 antibody can be paired with the purified BVD2-23B6 antibody (Cat. No. 554502) as the capture antibody, with recombinant human GM-CSF (Cat. No. 550068) as the standard. This detecting antibody preparation should be titrated from 0.5 - 2.0 µg/ml to determine its optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of GM-CSF protein ranging from ~15 to 2,000 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, ww.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook. 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; 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 OptEIA™ ELISA Set (Cat. No. 555126) is recommended. IF/Flow: The BVD2-21C11 antibody useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate GM-CSF producing cells within mixed cell populations. The PE-conjugated BVD2-21C11 antibody (Cat. No. 554507) is especially suitable for these studies. We also offer Alexa Fluor® 647 (Cat. No. 5622570) and PerCp-Cy™5.5 (Cat. No. 562258) formats. IP/WB: The BVD2-21C11 antibody has been reported to be useful for immunoprecipitation studies. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.
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.
