BV650 Rat Anti-Human IL-2

This is a suggested alternate SKU for 563467. Contact us for additional support.

BD Horizon™ BV650 Rat Anti-Human IL-2

Name: BV650 Rat Anti-Human IL-2
Brand: BD Horizon™
SKU: 564166

Clone MQ1-17H12

(RUO)

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™ BV650 Rat IgG2a, κ Isotype Control (Cat No. 563236; Left Panel) or BD Horizon BV650 Rat Anti-Human IL-2 antibody (Cat No. 564166; 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.

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Product Details

Brand: BD Horizon™

Alternative Name: IL2; Interleukin-2; T-cell growth factor; TCGF

Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)

Isotype: Rat IgG2a, κ

Immunogen: Human IL-2 Recombinant Protein

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

Entrez Gene ID: 3558

RRID: AB_2738637

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 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

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).
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.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
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.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

Fixation and Permeabilization Solution RUO

Size 125 mL Cat No. 554722

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

Protein Transport Inhibitor (Containing Monensin) RUO

Size 0.7 mL Cat No. 554724

View Details

564166 Rev. 2

Antibody Details

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™ BV650 which is part of the BD Horizon Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

564166 Rev. 2

Format Details

BV650

The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV650

Excitation Source: Violet 405 nm

Excitation Max: 406 nm

Emission Max: 649 nm

564166 Rev.2

Citations & References

Development References (3)

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
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
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

564166 Rev. 2

Please refer to Support Documents for Quality Certificates

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.