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PE Rat IgG2b, κ Isotype Control
PE Rat IgG2b, κ Isotype Control

Expression of IL-10 by MiCK-2 Cytokine Positive Control Cells. MiCK-2 cells (Cat. No. 554653) were permeabilized, and subsequently stained with 0.12 µg of PE-conjugated rat anti-mouse IL-10 antibody (PE-JES5-16E3, Cat. No. 554467; right panel) or 0.12 µg of PE-A95-1 rat IgG2b isotype control immunoglobulin (Cat. No. 556925; left panel) by using the BD Pharmingen staining protocol. The quadrant markers for the bivariate dot plots were set based on autofluorescence control.

Expression of IL-10 by MiCK-2 Cytokine Positive Control Cells. MiCK-2 cells (Cat. No. 554653) were permeabilized, and subsequently stained with 0.12 µg of PE-conjugated rat anti-mouse IL-10 antibody (PE-JES5-16E3, Cat. No. 554467; right panel) or 0.12 µg of PE-A95-1 rat IgG2b isotype control immunoglobulin (Cat. No. 556925; left panel) by using the BD Pharmingen staining protocol. The quadrant markers for the bivariate dot plots were set based on autofluorescence control.

製品詳細
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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_479625
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


調製と保管

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

推奨アッセイ手順

Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated A95-1 immunoglobulin (Cat. No. 556925) is a suitable rat IgG2b, κ isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse and human cells for flow cytometric analysis (see right panel). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

製品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. 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.
556925 Rev. 2
抗体の詳細
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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.

556925 Rev. 2
フォーマットの詳細
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
556925 Rev.2
引用&参考文献
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開発者向け参考資料 (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). 参考文献を見る
556925 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.