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FITC Mouse IgG2b, κ Isotype Control
FITC Mouse IgG2b, κ Isotype Control
Expression of human IL-8 by stimulated CD14+ human monocytes. Human PBMCs were stimulated for 6 hours with LPS (1.0 µg/ml final concentration; Sigma, Cat. No.L-8274) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). PBMCs were then harvested, stained with 0.25 µg of PE Mouse Anti-Human CD14 ( Cat. No. 555398), fixed, permeabilized, and subsequently stained with either 0.125 µg of FITC-anti-human IL-8 (left panel), or 0.125 µg of FITC Mouse IgG2b, κ Isotype Control (Cat. No. 555057; middle panel). The data reflect gating on monocytes, based on forward and side scattered light signals. The quadrant markers for the bivariate dot plot were set based on autofluorescence controls (right panel).
Expression of human IL-8 by stimulated CD14+ human monocytes. Human PBMCs were stimulated for 6 hours with LPS (1.0 µg/ml final concentration; Sigma, Cat. No.L-8274) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). PBMCs were then harvested, stained with 0.25 µg of PE Mouse Anti-Human CD14 ( Cat. No. 555398), fixed, permeabilized, and subsequently stained with either 0.125 µg of FITC-anti-human IL-8 (left panel), or 0.125 µg of FITC Mouse IgG2b, κ Isotype Control (Cat. No. 555057; middle panel). The data reflect gating on monocytes, based on forward and side scattered light signals. The quadrant markers for the bivariate dot plot were set based on autofluorescence controls (right panel).
Product Details
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BD Pharmingen™
Isotype Control (anti-Dansyl); Anti-Dansyl; Anti-DNS
Mouse C.SW IgG2b, κ
Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
0.5 mg/ml
AB_395677
Aqueous buffered solution containing protein stabilizer 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 FITC under optimum conditions, and unreacted FITC was removed.

Recommended Assay Procedures

Immunofluorescent Staining and Flow Cytometic Analysis: The PE-and FITC-conjugated 27-35 immunoglobulin (Cat. No. 555057, 555058 respectively) are suitable mouse IgG2b isotype controls for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized rat or human cells for flow cytometric analysis. Use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells) (see image, middle panel). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit our web site, www.bdbiosciences.com/us/s/resources and refer to the protocols section under Intracellular Flow.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
555057 Rev. 2
Antibody Details
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27-35

This mouse IgG2b, κ isotype control is a monoclonal antibody, clone 27-35, that is specific for the dansyl (5-[dimethylamino] naphthalene-1-sulfonyl) hapten. The dansyl (DNS) hapten is not expressed on human cells or human cell lines. The 27-35 immunoglobulin was selected as an isotype control following testing that demonstrated low background staining on a variety of mouse and human tissues.

555057 Rev. 2
Format Details
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FITC
Fluorescein (FITC) 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 518-nm. FITC 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.
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FITC
Blue 488 nm
494 nm
518 nm
555057 Rev.2
Citations & References
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Development References (3)

  1. BD Biosciences. Techniques for Immune Function Analysis, Application Handbook 1st Edition. 2003. Available: http://www.bdbiosciences.com/pdfs/manuals/02-8100055-21A1rr.pdf 2007, Jan. 25.
  2. Paliard P, Vitrey D, Fournier G, Belhadjali J, Patricot L, Berger F. Perhexiline maleate-induced hepatitis.. Digestion. 1978; 17(5):419-27. (Biology: Flow cytometry). 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). View Reference
555057 Rev. 2

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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.