Ly-76 / TER-119 Rat Monoclonal Antibody [Clone ID: TER-119]

CAT#: AM31858RP-N

Ly-76 / TER-119 rat monoclonal antibody, clone TER-119, PE

Product Images

Cell Source: Whole Blood. Percentage of cells stained above control: 98.0%.

Specifications

Product Data

• Clone Name: TER-119
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
  This Antibody has been reported to work in Western Blot, Immunoprecipitation and Immunohistochemistry on Frozen and Paraffin Sections.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Day 14 BALB/c fetal liver cells from Wistar Rat spleen.
• Specificity: Mouse Erythroid Cells (Ly-76).
  This anti-Mouse Erythroid cell (Ly-76) Monoclonal Antibody is selectively reactive with both fetal and adult erythroid cells.
  This monoclonal antibody (clone: TER119) is specific for cells at stages from early proerythroblast to mature erythrocytes.
  TER119 is reported to react with 20-25% of bone marrow cells and 2-3% of spleen cells but not with thymocytes or lymph node cells. In fetal haematopoietic tissues, 30-40% of day 10 yolk sac cells, 80-90% of day 14 fetal liver cells and 40-50% of newborn liver cells were reactive with AM31858RP. TER119+ cells in adult bone marrow expressed significant levels of CD45 but not myeloid (Mac-1, Gr-1) or B cell (B220) markers.
  This Monoclonal Antibody immunoprecipitated protein bands with molecular weights of 110 kDa, 60 kDa, 52 kDa and 32 kDa from erythrocyte membrane whereas only a 52 kDa band was detected by TER119 in Western Blot analysis. It has been determined that the TER119 antigen is a molecule associated with cell-surface glycophorin A but not with glycophorin A itself. Also the antigen is only expressed on normal erythroid cells but not on erythroleukaemia cells.
• Formulation: PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid purified Ig fraction.
• Concentration: 0.1 mg/ml
• Purification: Protein G Affinity Chromatography.
• Conjugation: PE
• Synonyms: Lymphocyte antigen 76, TER119
• Note: Test Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration: 1 x 10e6 cells per test
  Antibody Concentration Used: 0.05 μg/10e6 cells
  Secondary Antibody Used: PE Rat IgG2b
  
  Cell Source (Percentage of cells stained above control):
  Whole Bone Marrow (46.9%)
  Whole Blood (98.0%)
  *Blood was collected 1:1 in Alsever's and 0.1M Disodium EDTA was added 1:1 and incubated 10 minutes at room temperature followed by 3 washes with PBS
  
  Protocol: Flow Cytometry Analysis:
  
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population 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 this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test).
  4. To each tube, add 0.1-0.05 μg* of AM31858RP-N or AM31858RP-L 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.
  (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.)
  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 propidium iodide 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 (100 μl of 2M sodium azide in 100 mls).