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      Alexa Fluor® 488 Mouse IgG1 κ Isotype Control
      Alexa Fluor® 488 Mouse IgG1 κ Isotype Control
      Expression of TNF by stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells 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 Brefeldin A (GolgiPlug, Cat. No. 555029), Cells were harvested, fixed, permeabilized and stained with PE-conjugated mouse anti-human CD8 (PE-RPA-T8, Cat. No. 555367) and either mouse anti-human TNF antibody (Alexa Fluor® 488-MAb11, Cat. No. 557722), (left panel) or Alexa Fluor® 488-MOPC-21 immunoglobulin (Cat. No. 557721) (right panel) by using the staining protocol found in the protocols section at www.bdbioscience.com. To deomonstrate specificty of staining the binding of Alexa Fluor® 488-MAb11 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human TNF (Cat. No. 554618) and by preincubation of the fixed permeabilized cells with an excess of unlabelled MAb11 antibody (5 µg, Cat. No. 554510) prior to staining. 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.
      Alexa Fluor® 488 Mouse IgG1 κ Isotype Control
      Expression of TNF by stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells 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 Brefeldin A (GolgiPlug, Cat. No. 555029), Cells were harvested, fixed, permeabilized and stained with PE-conjugated mouse anti-human CD8 (PE-RPA-T8, Cat. No. 555367) and either mouse anti-human TNF antibody (Alexa Fluor® 488-MAb11, Cat. No. 557722), (left panel) or Alexa Fluor® 488-MOPC-21 immunoglobulin (Cat. No. 557721) (right panel) by using the staining protocol found in the protocols section at www.bdbioscience.com. To deomonstrate specificty of staining the binding of Alexa Fluor® 488-MAb11 was blocked by the preincubation of the conjugated antibody with molar excess of recombinant human TNF (Cat. No. 554618) and by preincubation of the fixed permeabilized cells with an excess of unlabelled MAb11 antibody (5 µg, Cat. No. 554510) prior to staining. 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™
      Mouse IgG1, κ
      Flow cytometry, Isotype control (Routinely Tested), Intracellular staining (flow cytometry) (Tested During Development)
      5 µl
      AB_396830
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      Recommended Assay Procedure:

      Immunofluorescent Staining and Flow Cytometric Analysis: The FITC-, PE-, APC-, PE-Cy7-, Alexa Fluor® 488-, and Alexa Fluor® 647-MOPC-21 immunoglobulins (Cat. No. 554679, 559320, 554681, 557646, 557721, and 557732) are suitable mouse IgG1κ isotype controls for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized mouse or human cells for flow cytometric analysis.

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      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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
      6. 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.
      7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
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      557702 Rev. 4
      Antibody Details
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      MOPC-21

      The MOPC-21 immunoglobulin is a mouse myeloma protein. The MOPC-21 immunoglobulin was selected as an isotype control following screening for low background on a variety of mouse and human tissues.

      Cat. no. 557721 has been optimized for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized mouse or human cells for flow cytometric analysis.  This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

      557702 Rev. 4
      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
      557702 Rev.4
      Citations & References
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      View product citations for antibody "557702" 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: Flow cytometry). View Reference
      557702 Rev. 4

      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.