This mAb reacts with T cells from mouse strains expressing the Ly 1.2 phenotype, but does not react with lymphocytes from mouse strains expressing the Ly 1.1 phenotype.
Purification
Protein G Chromatography
Immunogen
C3H.CE - Ly 1.2 : DS from C3H spleen. Fusion Partner: Myeloma SP2/0 - Ag 14 (M5).
CD5
Reactivity: Human
FACS
Host: Mouse
Monoclonal
L17F12
PE
Application Notes
Flow cytometry analysis (see Protocols). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 1 - 0. 2 µg* of this Ab per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 µl ice cold media B. 9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue DistributionMouse Strain: BALB/cCell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 0. 2 µg/10e6 cellsIsotypic Control: FITC Mouse IgG2b
Restrictions
For Research Use only
Concentration
0.1 mg/mL
Buffer
PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
CD5 is a 55 kDa T lymphocyte single chain transmembrane glycoprotein. It is present on all mature T lymphocytes, on most thymocytes and on many T cell leukemias and lymphomas. It reacts with a subpopulation of activated B cells. CD5/Lyt1 antigen is a monomeric type I transmembrane glycoprotein expressed on thymocytes, T lymphocytes, and a subset of B lymphocytes, but not on natural killer (NK) cells. It has been identified as the major ligand of the B cell antigen CD72. The frequency of CD5+ B cells exhibits strain dependent variation, and the phenotypic, anatomical, functional, developmental, and pathological characteristics of the CD5+ B cells suggest that they may represent a distinct lineage, known as B1 cells. Binding of CD5 on the T cell surface can augment alloantigen or mitogen induced lymphocyte proliferation and induces increased cytosolic free calcium, IL2 secretion, and IL2R expression. It has been proposed that CD5 negatively regulates signal transduction mediated by the T cell and B cell receptors.Synonyms: CD5, LEU1, Lymphocyte antigen T1/Leu-1, T-cell surface glycoprotein CD5