Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
China (Chinese)
登录/注册 UserName
产品

Link Arrow
解决方案

Link Arrow
发现&学习

Link Arrow
资源&工具

Link Arrow
Support

Link Arrow
Search icon Search icon
Close Menu
      Alert Icon
      PE Mouse IgG1, κ Isotype Control
      PE Mouse IgG1, κ Isotype Control
      Expression of IFN-γ by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC  were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. No. P-8139) and calcium ionophore A23187 (Sigma, Cat. No. C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy5 anti-CD3 (PE-Cy5-UCHT1, Cat. No. 555334), fixed, permeabilized, and subsequently stained with 0.25 µg of PE-mouse anti-human IFN-γ antibody (PE-4S.B3, Cat. No. 554552, left panel) or with 0.25 µg PE-MOPC-21 immunoglobulin (Cat. No. 554680, right panel) using the BD Pharmingen™ staining protocol. To demonstrate specificity of staining, the binding of PE-4S.B3 antibody was blocked by preincubation of fixed/permeabilized cells with excess unlabelled 4S.B3 antibody (5 µg; Cat. No. 554549). The quadrant markers for the bivariate dot plot were set based on autofluorescence controls and verified using the unlabelled 4S.B3 antibody blocking control.
      PE Mouse IgG1, κ Isotype Control
      Expression of IFN-γ by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC  were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. No. P-8139) and calcium ionophore A23187 (Sigma, Cat. No. C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy5 anti-CD3 (PE-Cy5-UCHT1, Cat. No. 555334), fixed, permeabilized, and subsequently stained with 0.25 µg of PE-mouse anti-human IFN-γ antibody (PE-4S.B3, Cat. No. 554552, left panel) or with 0.25 µg PE-MOPC-21 immunoglobulin (Cat. No. 554680, right panel) using the BD Pharmingen™ staining protocol. To demonstrate specificity of staining, the binding of PE-4S.B3 antibody was blocked by preincubation of fixed/permeabilized cells with excess unlabelled 4S.B3 antibody (5 µg; Cat. No. 554549). The quadrant markers for the bivariate dot plot were set based on autofluorescence controls and verified using the unlabelled 4S.B3 antibody blocking control.
      商品详情
      Down Arrow Up Arrow


      BD Pharmingen™
      Mouse IgG1, κ
      Flow cytometry, Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
      0.2 mg/ml
      AB_395506
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      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.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      推荐的实验流程

      Immunofluorescent Staining and Flow Cytometric Analysis: The PE-MOPC-21 immunoglobulins (Cat. No. 554680) is a suitable mouse IgG1κ isotype control for assessing the level of background staining on paraformaldehyde fixed/saponin-permeabilized mouse 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, right panel). For specific methodology, visit the protocols section of our website, or the chapter on intracellular staining in the Immune Function Handbook, which is posted on our web site at www.bdbiosciences.com. The intracellular cytokine staining technique and the use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      商品通知

      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.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow
      554680 Rev. 2
      抗体详情
      Down Arrow Up Arrow
      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.

      554680 Rev. 2
      格式详情
      Down Arrow Up Arrow
      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
      554680 Rev.2
      报价单和参考
      Down Arrow Up Arrow
      View product citations for antibody "554680" on CiteAb

      研发参考 (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. (Biology: Flow cytometry). 查看参考
      554680 Rev. 2

      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.