Alexa Fluor™ 488 Mouse Anti-Human IL-2

BD Pharmingen™ Alexa Fluor™ 488 Mouse Anti-Human IL-2

Name: Alexa Fluor™ 488 Mouse Anti-Human IL-2
Brand: Alexa Fluor™ 488 Mouse Anti-Human IL-2
SKU: 570115

Clone 5344.111 (also known as 5344) (RUO)

Two-color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml, 5 hr) and Calcium Ionophore A23187 (Sigma C-9275; 1 μg/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The permeabilized cells were then stained in BD Perm/Wash™ Buffer with BD Horizon™ BV786 Mouse Anti-Human CD3 antibody (Cat. No. 565491) and with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 567121; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human IL-2 antibody (Cat. No. 570034/570115; Right Plot). The pseudocolor density plot showing the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.

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Product Details

Brand: BD Pharmingen™

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

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human IL-2 protein

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

Vol. Per Test: 5 µl/test

Entrez Gene ID: 3558

RRID: AB_3685472

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

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
Alexa Fluor™ is a trademark of Life Technologies Corporation.
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.
For U.S. patents that may apply, see bd.com/patents.
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
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).

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

BV786 Mouse Anti-Human CD3 RUO

Size 50 Tests Cat No. 565491

Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control RUO

Size 0.1 mg Cat No. 567121

View Details

570115 Rev. 1

Antibody Details

5344.111

The 5344.111 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 and differentiation of many different target cell types including T cells, B cells, NK cells, monocytes and macrophages. The immunogen used to generate the 5344.111 hybridoma was affinity-purified, recombinant human IL-2 protein.

570115 Rev. 1

Format Details

Alexa Fluor™ 488

Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: Alexa Fluor™ 488

Excitation Source: Blue 488 nm

Excitation Max: 494 nm

Emission Max: 517 nm

570115 Rev.1

Citations & References

View product citations for antibody "570115" on CiteAb

Development References (7)

Bitmansour AD, Douek DC, Maino VC, Picker LJ. Direct ex vivo analysis of human CD4(+) memory T cell activation requirements at the single clonotype level.. J Immunol. 2002; 169(3):1207-18. (Clone-specific: Flow cytometry). View Reference
Jason J, Larned J. Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods. 1997; 207(1):13-22. (Clone-specific: Flow cytometry). View Reference
Liao W, Lin JX, Leonard WJ. IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Protoc Immunol. 2011; 23(5):598-604. (Biology). View Reference
Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
Mendes R, Bromelow KV, Westby M. Flow cytometric visualisation of cytokine production by CD3-CD56+ NK cells and CD3+CD56+ NK-T cells in whole blood. Cytometry. 2000; 39(1):72-78. (Clone-specific: Flow cytometry). 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). View Reference
Waldmann TA. The biology of interleukin-2 and interleukin-15: implications for cancer therapy and vaccine design. Nat Rev Immunol. 2006; 6(8):595-601. (Biology). View Reference

View All (7)

570115 Rev. 1

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.