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Preparation And Storage
Recommended Assay Procedures
Immunofluorescent Staining and Flow Cytometric Analysis: The APC-conjugated MQ1-17H12 antibody (Cat. No. 554567) can be used for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-2-producing cells within mixed cell populations (see image). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/1X10^6 cells). For specific methodology, please visit the protocols section under "Cytokines (Intracellular Staining)" on our web
site, http://www.bdbiosciences.com/us/s/resources.
A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MQ1-17H12 antibody with a molar excess of ligand (e.g., recombinant human IL-2; Cat No. 554603) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled MQ1-17H12 antibody (Cat. No. 554563) prior to staining. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is APC-R35-95 (Cat. No. 554690); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1X10^6 cells).
Neutralization/Blocking: The NA/LE™ format of the MQ1-17H12 antibody (Cat. No. 554567) is useful for neutralization of human IL-2 bioactivity. A suitable NA/LE™ rat IgG2a isotype control to match the NA/LE™ MQ1-17H12 antibody is the R35-95 antibody, Cat. No. 554687.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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The MQ1-17H12 monoclonal antibody specifically binds to the multifunctional cytokine, human Interleukin-2 (IL-2). IL-2 is produced by activated T cells and has multiple functions that can affect the growth, proliferation, differentiation and survival of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the MQ1-17H12 hybridoma was purified recombinant human IL-2 protein. The MQ1-17H12 antibody reportedly neutralizes the biological activity of human IL-2.
Development References (2)
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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. (Biology). 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. (Biology). View Reference
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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.