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Flow cytometric analysis of CD121a expression by mouse EL4.IL-2 cells. Cells from the mouse EL4.IL-2 (T lymphoma, ATCC TIB-181) cell line were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; dashed line histogram) or BD Horizon BV421 Rat Anti-Mouse CD121a antibody (Cat. No. 564387; solid line histogram). The fluorescence histogram showing CD121a expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable EL4.IL2 cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.


BD Horizon™ BV421 Rat Anti-Mouse CD121a

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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
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The 35F5 monoclonal antibody specifically binds to the Type I Interleukin-1 Receptor (IL-1 RI) which is also known as CD121a. CD121a is expressed by T-cell, fibroblast, and epithelial cell lines such as EL-4 and 3T3. Expression of the 35F5 epitope on normal mouse thymocytes and splenocytes is undetectable by flow cytometry. CD121a is an 80 kDa type-I transmembrane glycoprotein, a member of the Ig superfamily that mediates the inflammatory responses of leucocytes to IL-1. The 35F5 antibody is capable of blocking the binding of IL-1α, IL-1β, and IL-1 receptor antagonist to the Type I IL-1 Receptor. The 35F5 mAb is highly specific for the Type I IL-1 Receptor and does not react with the Type II IL-1 Receptor (CD121b) which is present on mouse macrophage and B-cell lines and polymorphonuclear leukocytes (PMN) and PMN progenitors.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

Development References (3)
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Chizzonite R, Truitt T, Kilian PL, et al. Two high-affinity interleukin 1 receptors represent separate gene products. Proc Natl Acad Sci U S A. 1989; 86(20):8029-8033. (Immunogen: Immunoprecipitation). View Reference
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McIntyre KW, Stepan GJ, Kolinsky KD, et al. Inhibition of interleukin 1 (IL-1) binding and bioactivity in vitro and modulation of acute inflammation in vivo by IL-1 receptor antagonist and anti-IL-1 receptor monoclonal antibody. J Exp Med. 1991; 173(4):931-939. (Clone-specific: Blocking). View Reference
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Zola H. Detection of cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.