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Alexa Fluor™ 647 Mouse Anti-Human IL-1β
Alexa Fluor™ 647 Mouse Anti-Human IL-1β

Flow cytometric analysis of IL-1β expression in activated human monocytes. Human peripheral blood mononuclear cells were cultured with Recombinant Human IFN-γ protein (Cat. No. 554616/554617; 10 ng/ml for 2 h at 37°C) and then stimulated with lipopolysaccharide (1 µg of LPS/ml) and BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (2 µM; Cat. No. 554724) overnight at 37°C. Cells were harvested, washed, and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Alexa Fluor™ 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732; dashed line histogram) or Alexa Fluor™ 647 Mouse Anti-Human IL-1β antibody (Cat. No. 567773/567774; solid line histogram). The fluorescence histogram showing IL-1β expression (or Ig Isotype control staining) was derived from gated events with the side and forward light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Flow cytometric analysis of IL-1β expression in activated human monocytes. Human peripheral blood mononuclear cells were cultured with Recombinant Human IFN-γ protein (Cat. No. 554616/554617; 10 ng/ml for 2 h at 37°C) and then stimulated with lipopolysaccharide (1 µg of LPS/ml) and BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (2 µM; Cat. No. 554724) overnight at 37°C. Cells were harvested, washed, and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Alexa Fluor™ 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732; dashed line histogram) or Alexa Fluor™ 647 Mouse Anti-Human IL-1β antibody (Cat. No. 567773/567774; solid line histogram). The fluorescence histogram showing IL-1β expression (or Ig Isotype control staining) was derived from gated events with the side and forward light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

商品详情
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BD Pharmingen™
IL-1 beta
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human IL-1β
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


准备和存储

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 to the dye under optimum conditions and unreacted dye was removed.

推荐的实验流程

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

商品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  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. 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).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
567773 Rev. 1
抗体详情
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AS10

The AS10 antibody reacts with human interleukin-1β (IL-1β) which is also known as endogenous pyrogen (EP), leukocyte endogenous mediator (LEM), mononuclear cell factor (MCF) and lymphocyte-activating factor (LAF). IL-1β is a proinflammatory cytokine that is synthesized as a precursor of 31 kDa and is converted intracellularly to the mature 17.5 kDa form, after cleavage by the IL-1β-converting enzyme (ICE). In healthy individuals, IL-1β is secreted non-constitutively by blood monocytes, tissue macrophages and dendritic cells. IL-1β is also constitutively expressed in the human hypothalamus. Many malignant tumors express IL-1β as part of their neoplastic nature.  The AS10 antibody has been reported not to recognize human IL-1α nor cross react with mouse IL-1β.

        

567773 Rev. 1
格式详情
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567773 Rev.1
报价单和参考
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研发参考 (5)

  1. Dinarello CA. Biology of interleukin 1. FASEB J. 1988; 2:108-115. (Biology).
  2. Dinarello CA. Interleukin-1 and interleukin-1 antagonism. Blood. 1991; 77(8):1627-1652. (Biology). 查看参考
  3. Mantovani A, Dejana E. Cytokines as communication signals between leukocytes and endothelial cells.. Immunol Today. 1989; 10(11):370-5. (Biology). 查看参考
  4. Oh KS, Gottschalk RA, Lounsbury NW, et al. Dual Roles for Ikaros in Regulation of Macrophage Chromatin State and Inflammatory Gene Expression. J Immunol. 2018; 201(2):757-771. (Clone-specific: Flow cytometry). 查看参考
  5. Slack J, McMahan CJ, Waugh S. et al. Independent binding of interleukin-1 α and interleukin-1 β to type I and type II interleukin 1 receptors. J Bio Chem. 1993; 268:2513-2524. (Biology).
查看所有文件 (5) 查看更少内容
567773 Rev. 1

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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.