Preparation And Storage
Recommended Assay Procedures
Flow cytometry: Since this antigen may be expressed at low density on the cell surface, investigators may wish to perform staining using Biotin Goat Anti-Rat Ig (Cat. No. 554014) coupled with a "bright" third-step reagent, such as PE Streptavidin (Cat. No. 554061).
Western Blot: The 35F5 antibody has been reported to be useful for western blotting of non-reduced proteins and being non-reactive to reduced proteins.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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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.
Development References (4)
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Callard R, Gearing A. Callard R, Gearing A. The Cytokine Facts Book. San Diego: Academic Press; 1994.
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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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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.