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PE Mouse Anti-Human IL-1β
PE 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 PE Mouse IgG1 κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human IL-1β antibody (Cat. No. 567779/567780; 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 PE Mouse IgG1 κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-Human IL-1β antibody (Cat. No. 567779/567780; 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.

Product Details
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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


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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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Antibody Details
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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.

        

567779 Rev. 1
Format Details
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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
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Citations & References
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Development References (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). View Reference
  3. Mantovani A, Dejana E. Cytokines as communication signals between leukocytes and endothelial cells.. Immunol Today. 1989; 10(11):370-5. (Biology). View Reference
  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). View Reference
  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).
View All (5) View Less
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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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.