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Purified Rat Anti-Mouse CD253
Product Details
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BD Pharmingen™
TRAIL; APO-2L; Ly-81; TL2
Mouse (QC Testing)
Rat F344, also known as Fischer, CDF IgG2a, κ
Mouse Trail-transfected 2PK-3 cells
Flow cytometry (Routinely Tested), Western blot (Reported)
0.5 mg/ml
AB_393605
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Since this antigen is expressed at low density on lymphoid cell surfaces, it may be desirable to amplify staining by using a biotinylated second-step antibody, such as goat anti-rat Ig polyclonal antibody (Cat. No. 554014), followed by a "bright" third-step reagent, such as Streptavidin-PE (Cat. No. 554061) or Streptavidin-APC (Cat. No. 554067). In order to obtain bright staining on LAK (Lymphokine-activated killer) cells, we suggested the use of A-LAK cells (adherent-LAK cells) that have been activated with 3,000 U/ml of recombinant mouse IL-2 (Cat. No. 550069) or human IL-2 (Cat. No. 554603). Other reported applications include western blot analysis and blocking of IL-2/IL-15-activated NK-cell cytotoxicity against mouse fibrosarcoma L929 target cells.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
550320 Rev. 6
Antibody Details
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N2B2

The N2B2 antibody reacts with mouse TRAIL (TNF-Related Apoptosis-Inducing Ligand, also known as APO-2 ligand or Ly-81), a 40-kDa type-II transmembrane protein of the TNF family. This antibody recognizes mouse Trail-transfected 2PK-3 cells, but not the parental cell line. TRAIL selectively induces apoptosis in various transformed cells, but not in normal cells. Five receptors have been identified for human TRAIL, whereas only one receptor has been described for the mouse counterpart thus far. The pattern of expression of mouse TRAIL is controversial. TRAIL has been detected, with a rabbit polyclonal antibody, on the surface of activated CD4+ and CD8+ T lymphocytes, as well as on CD45R/B220+ lymphocytes, and transformed T- and B-cell lines. Similar results were obtained by western blot and mRNA analyses. However, these results could not be reproduced with the N2B2 mAb. TRAIL expression, detected with the N2B2 mAb, is induced on CD3- NK1.1+ cells after stimulation with IL-2 or IL-15. However, neither freshly isolated nor activated T cells showed TRAIL expression on the cell surface. Furthermore, TRAIL mediates NK-cell cytotoxicity, which is blocked by the N2B2 mAb. TRAIL has been renamed as CD253 recently.

550320 Rev. 6
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
550320 Rev.6
Citations & References
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Development References (6)

  1. Degli-Esposti M. To die or not to die—the quest of the TRAIL receptors. J Leukoc Biol. 1999; 65:535-542. (Biology).
  2. Kayagaki N, Yamaguchi N, Nakayama M, et al. Expression and function of TNF-related apoptosis-inducing ligand on murine activated NK cells. J Immunol. 1999; 163(4):1906-1913. (Immunogen). View Reference
  3. Mariani SM, Krammer PH. Differential regulation of TRAIL and CD95 ligand in transformed cells of the T and B lymphocyte lineage. Eur J Immunol. 1998; 28(3):973-982. (Biology). View Reference
  4. Mariani SM, Krammer PH. Surface expression of TRAIL/Apo-2 ligand in activated mouse T and B cells. Eur J Immunol. 1998; 28(5):1492-1498. (Biology). View Reference
  5. Musgrave BL, Phu T, Butler JJ, Makrigiannis AP, Hoskin DW. Murine TRAIL (TNF-related apoptosis inducing ligand) expression induced by T cell activation is blocked by rapamycin, cyclosporin A, and inhibitors of phosphatidylinositol 3-kinase, protein kinase C, and protein tyrosine kinases: evidence for TRAIL induction via the T cell receptor signaling pathway. Exp Cell Res. 1999; 252(1):96-103. (Biology). View Reference
  6. Wu GS, Burns TF, Zhan Y, Alnemri ES, El-Deiry WS. Molecular cloning and functional analysis of the mouse homologue of the KILLER/DR5 tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor. Cancer Res. 1999; 59:2770-2775. (Biology).
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550320 Rev. 6

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