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PE Rat Anti-Mouse CD355 (CRTAM)
PE Rat Anti-Mouse CD355 (CRTAM)

Two-color flow cytometric analysis of CD355 (CRTAM) expression on mouse spleen cells. Splenic leucocytes from a C57BL/6 mouse were stimulated overnight with immobilized Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 µg/ml for plate coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 1 µg/ml) antibodies. The cells were harvested, washed, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and then stained with PerCP-Cy™5.5 Rat Anti-Mouse CD8a (Cat. No.551162) antibody and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD355 (CRTAM) antibody (Cat. No. 566748; Right Plot) at 0.5 µg/test. The pseudocolor density plot showing the correlated expression of CD355 (CRTAM) [or Ig Isotype control staining] versus PerCP-Cy™5.5 CD8a was derived from gated events with the forward and side light-scatter characteristics of viable activated leucocytes. Flow cytometry and data analysis were performed using a BD FACSCanto™ II System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of CD355 (CRTAM) expression on mouse spleen cells. Splenic leucocytes from a C57BL/6 mouse were stimulated overnight with immobilized Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 µg/ml for plate coating) and soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 1 µg/ml) antibodies. The cells were harvested, washed, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and then stained with PerCP-Cy™5.5 Rat Anti-Mouse CD8a (Cat. No.551162) antibody and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD355 (CRTAM) antibody (Cat. No. 566748; Right Plot) at 0.5 µg/test. The pseudocolor density plot showing the correlated expression of CD355 (CRTAM) [or Ig Isotype control staining] versus PerCP-Cy™5.5 CD8a was derived from gated events with the forward and side light-scatter characteristics of viable activated leucocytes. Flow cytometry and data analysis were performed using a BD FACSCanto™ II System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
class I-restricted T cell-associated molecule; crtam; cytotoxic and regulatory T-cell molecule
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Mouse CRTAM extracellular sequence Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869846
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. 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. Cy is a trademark of GE Healthcare.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566748 Rev. 1
Antibody Details
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11-5

The 11-5 monoclonal antibody specifically recognizes MHC class I-restricted T cell-associated molecule (CRTAM) that is also known as CD355 or Cytotoxic and regulatory T-cell molecule. CD355 (CRTAM) is encoded by Crtam which belongs to the Nectin family of cell adhesion molecules within the immunoglobulin superfamily. This type I transmembrane glycoprotein contains a V-type Ig domain, followed by a C-type Ig domain, a transmembrane region, and an intracellular region that contains a PDZ-binding motif at the C terminus. CD355 (CRTAM) is transiently expressed as a homodimer on activated NK cells, NKT cells, γδ T cells, CD8+ T cells, or CD4+ T cells. Through its V-type Ig domain, CRTAM (CD355) binds to Nectin-like molecule 2 (Necl-2) and plays important roles in adhesive interactions between different cell types and in cellular migration.

        

566748 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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566748 Rev.1
Citations & References
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Development References (5)

  1. Arase N, Takeuchi A, Unno M, et al. Heterotypic interaction of CRTAM with Necl2 induces cell adhesion on activated NK cells and CD8+ T cells.. Int Immunol. 2005; 17(9):1227-37. (Immunogen: Flow cytometry). View Reference
  2. Fuchs A, Colonna M. The role of NK cell recognition of nectin and nectin-like proteins in tumor immunosurveillance. Semin Cancer Biol. 2006; 16(5):359-366. (Biology). View Reference
  3. Kennedy J, Vicari AP, Saylor V, et al. A molecular analysis of NKT cells: identification of a class-I restricted T cell-associated molecule (CRTAM).. J Leukoc Biol. 2000; 67(5):725-34. (Biology). View Reference
  4. Takeuchi A, Badr Mel S, Miyauchi K, et al. CRTAM determines the CD4+ cytotoxic T lymphocyte lineage. J Exp Med. 2016; 213(1):123-38. (Clone-specific: Flow cytometry). View Reference
  5. Takeuchi A, Itoh Y, Takumi A, et al. CRTAM confers late-stage activation of CD8+ T cells to regulate retention within lymph node. J Immunol. 2009; 183(7):4220-4228. (Biology). View Reference
View All (5) View Less
566748 Rev. 1

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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.