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BV421 Rat Anti-Human IL-2
BV421 Rat Anti-Human IL-2
Two color flow cytometric analysis of IL-2 expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 μ g/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were fixed with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722), washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723).        The cells were then stained with PE Mouse Anti-Human CD3 antibody (Cat No. 555333/561808/561809) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat No. 562602, Left Panel) or BD Horizon BV421 Rat Anti-Human IL-2 antibody (Cat No. 564164, Right Panel) using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric contour plots showing correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two color flow cytometric analysis of IL-2 expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 μ g/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were fixed with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722), washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723).        The cells were then stained with PE Mouse Anti-Human CD3 antibody (Cat No. 555333/561808/561809) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat No. 562602, Left Panel) or BD Horizon BV421 Rat Anti-Human IL-2 antibody (Cat No. 564164, Right Panel) using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric contour plots showing correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
IL2; Interleukin-2; T-cell growth factor; TCGF
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Rat IgG2a, κ
Human IL-2 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
3558
AB_2738635
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349).

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564164 Rev. 2
Antibody Details
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MQ1-17H12

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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

564164 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564164 Rev.2
Citations & References
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View product citations for antibody "564164" on CiteAb

Development References (3)

  1. 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
  2. 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: Blocking, ELISA, Immunoprecipitation). View Reference
  3. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Flow cytometry). View Reference
564164 Rev. 2

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.