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PE Mouse IgG2a, κ Isotype Control
PE Mouse IgG2a, κ Isotype Control

Expression of human MIP-1α by stimulated CD14+ human lymphocytes. Human PBMC were stimulated for 6 hours with LPS (100 ng/ml final concentration) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with FITC-mouse anti-human CD14 monoclonal antibody (FITC-M5E2, Cat. No. 555397), fixed permeabilized, and subsequently stained with either PE-anti-human MIP-1α (Cat. No. 554730, top panel), or PE-mouse IgG2aκ (Cat. No. 559319; middle panel), by following the Pharmingen staining protocols. The data reflect gating on monocytes, based on forward and scattered light signals. The quadrant markers for the bivariate dot plot were set based on autofluorescence controls (bottom panel).

Expression of human MIP-1α by stimulated CD14+ human lymphocytes. Human PBMC were stimulated for 6 hours with LPS (100 ng/ml final concentration) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with FITC-mouse anti-human CD14 monoclonal antibody (FITC-M5E2, Cat. No. 555397), fixed permeabilized, and subsequently stained with either PE-anti-human MIP-1α (Cat. No. 554730, top panel), or PE-mouse IgG2aκ (Cat. No. 559319; middle panel), by following the Pharmingen staining protocols. The data reflect gating on monocytes, based on forward and scattered light signals. The quadrant markers for the bivariate dot plot were set based on autofluorescence controls (bottom panel).

製品詳細
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BD Pharmingen™
Mouse BALB/c IgG2a, κ
TNP-keyhole limpet hemocyanin
Flow cytometry, Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
20 µl
AB_397217
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


調製と保管

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

推奨アッセイ手順

Immunofluorescent Staining and Flow Cytometric Analysis: The PE-conjugated G155-178 immunoglobulins are a suitable mouse IgG2a, κ isotype control for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized rat or human cells for flow cytometric analysis.  For specific methodology, please visit our web site, www.bdbiosciences.com and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook. The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

This pre-titered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized. Perm/Wash™ Buffer (Cat. No. 554723) contains the permeabilization agent saponin and is useful for this purpose.

製品通知

  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
559319 Rev. 2
抗体の詳細
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G155-178

The G155-178 clone has an unknown specificity. Trinitrophenol (TNP), the immunogen, is a hapten not expressed on human, mouse, rat or non-human primate cells. In the absence of specific binding, this antibody may bind non-specifically to immunoglobulin Fc receptors. The immunoglobulin secreted by the G155-178 hybridoma was selected as a mouse IgG2a, κ isotype control following screening for low background binding on a variety of mouse and human tissues.

559319 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
559319 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: Flow cytometry). 参考文献を見る
559319 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.