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Biotin Rat Anti-Mouse TIM-2
Biotin Rat Anti-Mouse TIM-2

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 Biotin Rat IgG2b, κ Isotype Control (Cat. No. 555847; dotted line histograms) or Biotin Rat Anti-Mouse TIM-2 antibody (Cat. No. 566592; solid line histograms) at 0.5 μg/test. The cells were then stained with PE Streptavidin (Cat. No. 554061) and 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.

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 Biotin Rat IgG2b, κ Isotype Control (Cat. No. 555847; dotted line histograms) or Biotin Rat Anti-Mouse TIM-2 antibody (Cat. No. 566592; solid line histograms) at 0.5 μg/test. The cells were then stained with PE Streptavidin (Cat. No. 554061) and 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
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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)
0.5 mg/ml
171284
AB_2739754
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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.
566592 Rev. 1
Antibody Details
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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.

        

566592 Rev. 1
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
566592 Rev.1
Citations & References
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Development References (8)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
View All (8) View Less
566592 Rev. 1

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