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Flow cytometric analysis of CD365 (TIM-1) expression on activated mouse B cells. BALB/c mouse splenic B cells were isolated with BD IMag™ Anti-Mouse CD45R/B220 Magnetic Particles-DM (Cat. No. 551513) and cultured for 3 days in media alone (Left Panel) or stimulated (Right Panel) with AffiniPure F(ab')2 Goat Anti-Mouse Anti-IgM (Jackson ImmunoResearch) and Purified NA/LE Hamster Anti-Mouse CD40 antibody (Cat. No. 553721). The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with either 0.25 μg of PE Rat IgG2b, κ Isotype Control (Cat. No. 556925; dashed line histograms) or PE Rat Anti-Mouse CD365 (TIM-1) antibody (Cat. No. 566335; solid line histograms). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Histograms showing CD365 (TIM-1) expression were derived from 7-AAD-negative gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse CD365 (TIM-1)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- An isotype control should be used at the same concentration as the antibody of interest.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The RMT1-4 monoclonal antibody specifically recognizes CD365 which is encoded by Havcr1 (Hepatitis A virus cellular receptor 1) and commonly known as T cell immunoglobulin and mucin domain-containing protein 1 (TIM-1). TIM-1 is a type I transmembrane glycoprotein that contains an extracellular immunoglobulin V-like domain and mucin-like domain and a conserved intracellular tyrosine phosphorylation motif that is involved in transmembrane signaling. TIM-1 is a costimulatory molecule that plays an important role in regulating immune responses. It is expressed on mast cells, and activated T cells including T helper type 2 (Th2) cells. TIM-1 is constitutively expressed by some B cells with increased TIM-1 expression detected upon B cell activation. TIM-1 was reportedly expressed by the majority of IL-10-expressing regulatory B cells within several major splenic B cell subsets. These cells may play critical roles in Th2-like CD4 T cell differentiation and in the regulation of inflammation and autoimmunity. TIM-4, a phosphatidylserine receptor encoded by Timd4, is a putative ligand for TIM-1.
Development References (6)
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Ding Q, Yeung M, Camirand G, et al. Regulatory B cells are identified by expression of TIM-1 and can be induced through TIM-1 ligation to promote tolerance in mice.. J Clin Invest. 2011; 121(9):3645-56. (Clone-specific: Flow cytometry). View Reference
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Kuchroo VK, Umetsu DT, DeKruyff RH, Freeman GJ. The TIM gene family: emerging roles in immunity and disease.. Nat Rev Immunol. 2003; 3(6):454-62. (Biology). View Reference
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Ma J, Usui Y, Takeda K, et al. TIM-1 signaling in B cells regulates antibody production.. Biochem Biophys Res Commun. 2011; 406(2):223-8. (Biology: Flow cytometry). View Reference
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Meyers JH, Chakravarti S, Schlesinger D, et al. TIM-4 is the ligand for TIM-1, and the TIM-1-TIM-4 interaction regulates T cell proliferation.. Nat Immunol. 2005; 6(5):455-64. (Biology). View Reference
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Nakae S, Iikura M, Suto H, et al. TIM-1 and TIM-3 enhancement of Th2 cytokine production by mast cells.. Blood. 2007; 110(7):2565-8. (Immunogen: Flow cytometry). View Reference
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Umetsu SE, Lee WL, McIntire JJ, et al. TIM-1 induces T cell activation and inhibits the development of peripheral tolerance.. Nat Immunol. 2005; 6(5):447-54. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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