Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
Australia (English)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Link Arrow
Solutions

Link Arrow
Discover & Learn

Link Arrow
Resources & Tools

Link Arrow
Support

Link Arrow
Search icon Search icon
Close Menu
      Alert Icon
      Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control

      BD Pharmingen™ Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control

      Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control
      Identification of activated T-lymphocytes using the Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control.  BALB/c splenocytes were activated in culture for 48 hours using plate-bound hamster anti-mouse CD3e antibody (clone 500A2) (Cat. No. 553238) and stained with either Mouse T Lymphocyte Activation Isotype Control (upper and lower left panels) or Mouse T Lymphocyte Activation Antibody Cocktail (upper and lower middle panels).  In addition, unactivated BALB/c splenocytes were stained with the Mouse T Lymphocyte Activation Antibody Cocktail (upper and lower right panels) or the Mouse T Lymphocyte Activation Isotype Control (not shown).  Scatter plots were used to select either activated lymphoblasts (left and middle panels) or resting/unactivated lymphocytes (right panels) for data analysis.  The two-color contour plots display the CD3e+ T lymphocytes which express the activation antigens CD25 (upper middle and right panels) and CD69 (lower middle and right panels).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry instrument.
      Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control
      Identification of activated T-lymphocytes using the Mouse T Lymphocyte Activation Antibody Cocktail, with Isotype Control.  BALB/c splenocytes were activated in culture for 48 hours using plate-bound hamster anti-mouse CD3e antibody (clone 500A2) (Cat. No. 553238) and stained with either Mouse T Lymphocyte Activation Isotype Control (upper and lower left panels) or Mouse T Lymphocyte Activation Antibody Cocktail (upper and lower middle panels).  In addition, unactivated BALB/c splenocytes were stained with the Mouse T Lymphocyte Activation Antibody Cocktail (upper and lower right panels) or the Mouse T Lymphocyte Activation Isotype Control (not shown).  Scatter plots were used to select either activated lymphoblasts (left and middle panels) or resting/unactivated lymphocytes (right panels) for data analysis.  The two-color contour plots display the CD3e+ T lymphocytes which express the activation antigens CD25 (upper middle and right panels) and CD69 (lower middle and right panels).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry instrument.
      Product Details
      Down Arrow Up Arrow


      BD Pharmingen™
      Mouse (QC Testing)
      Flow cytometry (Routinely Tested)
      RUO
      AB_396937


      Description

      The Mouse T Lymphocyte Activation Antibody Cocktail is a three-color reagent designed to identify major subsets of T lymphocytes by direct immunofluorscent staining using flow cytometric analysis.  This cocktail consists of the following antibody mixture:  PE-Cy™7 rat anti-mouse CD25 (clone PC61), PE Armenian hamster anti-mouse CD69 (clone H1.2F3), and FITC Armenian hamster anti-mouse CD3ε (clone 145-2C11).  PE-Cy™7 rat anti-mouse CD25 (clone PC61) reacts with CD25, the low-affinity IL-2 receptor α chain (IL-2Rα, p55) expressed on activated mouse T and B lymphocytes.  CD25 is also found on some developing B cells in the bone marrow, early developing T cells in the thymus, peripheral CD4+ regulatory T (Treg) cells, and dendritic cells.  PE hamster anti-mouse CD69 (clone H1.2F3) reacts with CD69, also known as the very early activation antigen.  Its expression is rapidly induced upon activation of lymphocytes (T, B, NK, and NK-T cells), neutrophils, and macrophages.  CD69 has also been reported to be expressed on thymocytes that are undergoing positive selection.  FITC hamster anti-mouse CD3ε (clone 145-2C11) reacts with the 25 kDa ε chain of the T-cell receptor-associated CD3 complex, expressed on thymocytes, mature T lymphocytes, and NK-T cells.

      The Mouse T Lymphocyte Activation Isotype Control contains equivalent concentrations of fluorochrome- and isotype-matched negative-control immunoglobulin consisting of the following: PE-Cy™7 rat IgG1, λ (clone A110-1), PE Armenian hamster IgG1, λ (clone G235-2356) and FITC Armenian hamster IgG1, κ (clone A19-3).

      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 FITC under optimum conditions, and unreacted FITC was removed. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. The antibody was conjugated with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
      Show More blue down arrow Show Less blue up arrow

      Recommended Assay Procedures

      Flow cytometry:  The three antibodies contained within the Mouse T Lymphocyte Activation Antibody Cocktail have been titrated, pre-diluted, mixed together, and formulated for optimal staining performance.  The use of three different fluorochromes for labeling of three different antibodies allows for the distinct recognition of three different antigens on each cell in a sample.  The levels of expression of the three antigens distinguish the major subpopulations of developing and peripheral T lymphocytes.  Additional fluorochrome-labeled reagents may be combined with the Mouse T Lymphocyte Activation Antibody Cocktail, and the Mouse T Lymphocyte Activation Isotype Control, to further characterize T-cell subpopulations.

      Show More blue down arrow Show Less blue up arrow

      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      6. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
      7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      8. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
      9. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      10. Cy is a trademark of GE Healthcare.
      11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      Show More blue down arrow Show Less blue up arrow
      557916 Rev. 3
      Components
      Down Arrow Up Arrow
      Description Quantity/Size Part Number EntrezGene ID
      Mouse T Lymphocyte Activation Antibody Cocktail 100 Tests (1 ea) 51-9002232 N/A
      Mouse T Lymphocyte Activation Isotype Control 100 Tests (1 ea) 51-9002243 N/A
      557916 Rev. 3
      Citations & References
      Down Arrow Up Arrow

      Development References (19)

      1. Bendelac A, Matzinger P, Seder RA, Paul WE, Schwartz RH. Activation events during thymic selection. J Exp Med. 1992; 175(3):731-742. (Biology). View Reference
      2. Brandle D, Muller S, Muller C, Hengartner H, Pircher H. Regulation of RAG-1 and CD69 expression in the thymus during positive and negative selection. Eur J Immunol. 1994; 24(1):145-151. (Biology). View Reference
      3. Ceredig R, Lowenthal JW, Nabholz M, MacDonald HR. Expression of interleukin-2 receptors as a differentiation marker on intrathymic stem cells. Nature. 1985; 314(6006):98-100. (Biology). View Reference
      4. Chen J, Ma A, Young F, Alt FW. IL-2 receptor alpha chain expression during early B lymphocyte differentiation. Int Immunol. 1994; 6(8):1265-1268. (Biology). View Reference
      5. Garni-Wagner BA, Witte PL, Tutt MM, et al. Natural killer cells in the thymus. Studies in mice with severe combined immune deficiency. J Immunol. 1990; 144(3):796-803. (Biology). View Reference
      6. Godfrey DI, Zlotnik A. Control points in early T-cell development. Immunol Today. 1993; 14(11):547-553. (Biology). View Reference
      7. Leo O, Foo M, Sachs DH, Samelson LE, Bluestone JA. Identification of a monoclonal antibody specific for a murine T3 polypeptide. Proc Natl Acad Sci U S A. 1987; 84(5):1374-1378. (Biology). View Reference
      8. Lowenthal JW, Corthésy P, Tougne C, Lees R, MacDonald HR, Nabholz M. High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61. J Immunol. 1985; 135(6):3988-3994. (Biology). View Reference
      9. Lowenthal JW, Zubler RH, Nabholz M, MacDonald HR. Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes. Nature. 1985; 315(6021):669-672. (Biology). View Reference
      10. Maruoka H, Ikarashi Y, Shinohara K, et al. A novel monoclonal antibody permitting recognition of NKT cells in various mouse strains. Biochem Biophys Res Commun. 1998; 242(2):413-418. (Biology). View Reference
      11. Marzio R, Jirillo E, Ransijn A, Mauel J, Corradin SB. Expression and function of the early activation antigen CD69 in murine macrophages. J Leukoc Biol. 1997; 62(3):349-355. (Biology). View Reference
      12. Nishimura T, Kitamura H, Iwakabe K, et al. The interface between innate and acquired immunity: glycolipid antigen presentation by CD1d-expressing dendritic cells to NKT cells induces the differentiation of antigen-specific cytotoxic T lymphocytes. Int Immunol. 2000; 12(7):987-994. (Biology). View Reference
      13. Read S, Malmstrom V, Powrie F. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation. J Exp Med. 2000; 192(2):295-302. (Biology). View Reference
      14. Rolink A, Grawunder U, Winkler TH, Karasuyama H, Melchers F. IL-2 receptor alpha chain (CD25, TAC) expression defines a crucial stage in pre-B cell development. Int Immunol. 1994; 6(8):1257-1264. (Biology). View Reference
      15. Takahashi T, Tagami T, Yamazaki S, et al. Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4. J Exp Med. 2000; 192(2):303-309. (Biology). View Reference
      16. Taniguchi T, Minami Y. The IL-2/IL-2 receptor system: a current overview. Cell. 1993; 73(1):5-8. (Biology). View Reference
      17. Yokoyama WM, Koning F, Kehn PJ, et al. Characterization of a cell surface-expressed disulfide-linked dimer involved in murine T cell activation. J Immunol. 1988; 141(2):369-376. (Biology). View Reference
      18. Yokoyama WM, Maxfield SR, Shevach EM. Very early (VEA) and very late (VLA) activation antigens have distinct functions in T lymphocyte activation. Immunol Rev. 1989; 109:153-176. (Biology). View Reference
      19. Ziegler SF, Ramsdell F, Alderson MR. The activation antigen CD69. Stem Cells. 1994; 12(5):456-465. (Biology). View Reference
      View All (19) View Less
      557916 Rev. 3

      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.

      Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.