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

Specifications

Product Data
Clone Name	TER-119
Applications	FC, IHC, IP
Recommend Dilution	Flow Cytometry (See Protocols). This Antibody has been reported to work in Western Blot, Immunoprecipitation and Immunohistochemistry on Frozen and Paraffin Sections.
Reactivity	Mouse
Host	Rat
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 AM31858PU-N. 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: Biotin State: Liquid purified Ig fraction.
Concentration	lot specific
Purification	Protein G Affinity Chromatography.
Conjugation	Biotin
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
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: Biotin Rat IgG2b Cell Source (Percentage of cells stained above control): Whole Bone Marrow (24.8%) Whole Blood (100%) *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 AM31858BT-N or AM31858BT-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. 7. Wash 2 times at 4°C. 8. Add 100 μl of secondary antibody (FITC-Streptavidin) at 1/500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 11. Resuspend the cell pellet in 50 μl ice cold media B. 12. 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).
Reference Data

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