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Alexa Fluor® 488 Rat IgG2b, κ Isotype Control
Alexa Fluor® 488 Rat IgG2b, κ Isotype Control
Expression of IL-2 by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA (5 ng/ml, Sigma, Cat. No. P-8139) and Ionomycin (500 ng, Sigma, P-8139) in the presence of BD GolgiPlug™ (Cat. No. 555029). Cells were harvested, fixed, permeabilized, and stained with PE-conjugated rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553048) and either rat anti-mouse IL-2 antibody (Alexa Fluor® 488-JES6-5H4, Cat. No. 557725), (left panel) or immunoglobulin isotype control (Alexa Fluor® 488-A95-1, Cat. No. 557726), (right panel) by using Pharmingen's staining protocol. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.
Expression of IL-2 by stimulated CD4+ and CD4- BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA (5 ng/ml, Sigma, Cat. No. P-8139) and Ionomycin (500 ng, Sigma, P-8139) in the presence of BD GolgiPlug™ (Cat. No. 555029). Cells were harvested, fixed, permeabilized, and stained with PE-conjugated rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553048) and either rat anti-mouse IL-2 antibody (Alexa Fluor® 488-JES6-5H4, Cat. No. 557725), (left panel) or immunoglobulin isotype control (Alexa Fluor® 488-A95-1, Cat. No. 557726), (right panel) by using Pharmingen's staining protocol. Dot plots were derived from gated events with the forward and side light scatter characteristics of lymphocytes. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.
Product Details
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BD Pharmingen™
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2b, κ
TNP-Keyhole Limpet Hemocyanin
Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
0.2 mg/ml
AB_396834
Aqueous buffered solution containing ≤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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  3. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
557726 Rev. 2
Antibody Details
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A95-1

The A95-1 antibody has unknown specificity. Trinitrophenal (TNP), the immunogen, is a hapten that is not expressed on human, mouse, rat, or non-human primate cells. The A95-1 immunoglobulin was selected as an isotype control following screening for low background on a variety of mouse and human tissues.

557726 Rev. 2
Format Details
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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.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
557726 Rev.2
Citations & References
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View product citations for antibody "557726" on CiteAb

Development References (1)

  1. 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
557726 Rev. 2

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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.