Flow cytometric analysis of TIM-2 expression on activated mouse B cells. BALB/c mouse splenocytes were stimulated in culture for 3 days with Purified NA/LE Hamster Anti-Mouse CD40 antibody (Cat. No. 553721) alone or in combination with either Recombinant Mouse IL-4 protein (Cat. No. 550067) or Donkey Anti-Mouse IgM F(ab')2 (Jackson ImmunoResearch Labs Cat. No. 715-006-020) as indicated. The cells were harvested, preincubated with BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with Alexa Fluor® 647 Rat Anti-CD45R/B220 antibody (Cat. No. 557683) and either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; dotted line histograms) or PE Rat Anti-Mouse TIM-2 antibody (Cat. No. 566591; solid line histograms) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The histograms showing TIM-2 expression (or Ig Isotype Control staining) were derived from CD45R/B220-positive and 7-AAD-negative gated events with the with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
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Flow cytometric analysis of TIM-2 expression on activated mouse B cells. BALB/c mouse splenocytes were stimulated in culture for 3 days with Purified NA/LE Hamster Anti-Mouse CD40 antibody (Cat. No. 553721) alone or in combination with either Recombinant Mouse IL-4 protein (Cat. No. 550067) or Donkey Anti-Mouse IgM F(ab')2 (Jackson ImmunoResearch Labs Cat. No. 715-006-020) as indicated. The cells were harvested, preincubated with BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with Alexa Fluor® 647 Rat Anti-CD45R/B220 antibody (Cat. No. 557683) and either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; dotted line histograms) or PE Rat Anti-Mouse TIM-2 antibody (Cat. No. 566591; solid line histograms) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The histograms showing TIM-2 expression (or Ig Isotype Control staining) were derived from CD45R/B220-positive and 7-AAD-negative gated events with the with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
T-cell membrane protein 2; Tim-2; Tim2; TIMD-2; Timd2
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Mouse TIM-2 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
171284
AB_2739753
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
- 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.
- 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
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
PE Rat IgG2b, κ Isotype Control RUO
Size
100 Tests
Cat No.
555848
Purified NA/LE Hamster Anti-Mouse CD40 RUO
Size
0.5 mg
Cat No.
553721
566591 Rev. 1
Antibody Details
RMT2-26
The RMT2-26 monoclonal antibody specifically recognizes T cell immunoglobulin and mucin domain-2 (TIM-2) which is also known as T-cell membrane protein 2, TIMD-2, or Tim2. TIM-2 is an ~55 kDa type I transmembrane glycoprotein that is encoded by Timd2 (T cell immunoglobulin and mucin domain containing 2) which belongs to the immunoglobulin gene superfamily. TIM-2 contains extracellular IgV and mucin domains and a cytoplasmic region with a conserved tyrosine phosphorylation motif which is involved in transmembrane signaling. A human ortholog for mouse TIM-2 has not been identified. TIM-2 is reportedly expressed on B cells with higher levels on activated B cells and germinal center B cells, type-2 T helper (Th2) cells, oligodendrocytes, and epithelial cells in the kidney and liver. It may serve as a receptor for Semaphorin 4A (Sema4A) or the heavy chain of ferritin (H-ferritin). The RMT2-26 antibody can reportedly block the binding of H-ferritin to TIM-2.
566591 Rev. 1
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566591 Rev.1
Citations & References
Development References (8)
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Abe Y, Kamachi F, Kawamoto T, et al. TIM-4 has dual function in the induction and effector phases of murine arthritis.. J Immunol. 2013; 191(9):4562-72. (Clone-specific: Flow cytometry). View Reference
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Chakravarti S, Sabatos CA, Xiao S, et al. Tim-2 regulates T helper type 2 responses and autoimmunity.. J Exp Med. 2005; 202(3):437-44. (Biology). View Reference
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Chen TT, Li L, Chung DH, et al. TIM-2 is expressed on B cells and in liver and kidney and is a receptor for H-ferritin endocytosis.. J Exp Med. 2005; 202(7):955-65. (Biology). View Reference
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Han J, Seaman WE, Di X, et al. Iron uptake mediated by binding of H-ferritin to the TIM-2 receptor in mouse cells.. PLoS ONE. 2011; 6(8):e23800. (Biology). View Reference
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Kawamoto T, Abe Y, Ito J, et al. Anti-T cell immunoglobulin and mucin domain-2 monoclonal antibody exacerbates collagen-induced arthritis by stimulating B cells.. Arthritis Res Ther. 2011; 13(2):R47. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Western blot). View Reference
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Kumanogoh A, Marukawa S, Suzuki K, et al.. Class IV semaphorin Sema4A enhances T-cell activation and interacts with Tim-2. Nature. 2002; 419(6907):629-633. (Biology). View Reference
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Rennert PD, Ichimura T, Sizing ID, et al. T cell, Ig domain, mucin domain-2 gene-deficient mice reveal a novel mechanism for the regulation of Th2 immune responses and airway inflammation.. J Immunol. 2006; 177(7):4311-21. (Biology). View Reference
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Rodriguez-Manzanet R, DeKruyff R, Kuchroo VK, Umetsu DT. The costimulatory role of TIM molecules. Immunol Rev. 2009; 229(1):259-270. (Biology). View Reference
566591 Rev. 1
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.
