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PE Rat Anti-Mouse CD365 (TIM-1)
PE Rat Anti-Mouse CD365 (TIM-1)

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.

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.

Product Details
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BD Pharmingen™
Havcr1; Tim1; TIMD-1; Timd1; KIM-1; Kidney injury molecule 1
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Mouse TIM-1 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739692
Aqueous buffered solution containing ≤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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. An isotype control should be used at the same concentration as the antibody of interest.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566335 Rev. 1
Antibody Details
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RMT1-4

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.

        

566335 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
566335 Rev.1
Citations & References
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Development References (6)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
View All (6) View Less
566335 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.