Thy1 Mouse Monoclonal Antibody [Clone ID: 5a-8]

CAT#: CL039APC

Thy1 mouse monoclonal antibody, clone 5a-8, APC

Product Images

Cell Source: Thymus Percentage of cells stained above control: 99.8%

Specifications

Product Data

• Clone Name: 5a-8
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: CBA/J.
  Donor: AKR/J Spleen.
  Fusion Partner: Spleen from immunized recipient fused with myeloma P3-NSI-1-Ag4-1.
• Specificity: This monoclonal antibody reacts with all T lymphocytes from mouse strains expressing the Thy 1.2 phenotype (e.g. C57BL/6, C3H/He, DBA/2, CBA/J, BALB/c), but does not react with lymphocytes expressing the Thy 1.1 phenotype [e.g. AKR/J, B6.PL(74NS)].
• 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: APC
  State: Liquid purified IgG fraction.
• Concentration: 0.2 mg/ml
• Conjugation: APC
• Gene Name: Mus musculus thymus cell antigen 1, theta (Thy1)
• Database Link:
  Entrez Gene 21838 Mouse
• Background: CD90 (Thy1) antigen is a GPI linked glycoprotein member of the Immunoglobulin superfamily. It is expressed on murine T cells, thymocytes, neural cells, cells of granulocytic lineage, early hematopoietic progenitors, fibroblasts, neurons and Kupffer's cells. Thy1 may play a role in cell to cell or cell to ligand interactions during synaptogenesis and other events in the brain. It is found in most mouse strains except AKR/J, A, Thy1.1 and B6.PL (74NS) expressing Thy1.1.
• Synonyms: Thy-1, THY1, CDw90
• Note: 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 1.0 µg* of this CL039APC 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).
  
  Results - Tissue Distribution:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Isotypic Control: APC Mouse IgG2b
  
  Cell Source Percentage of cells stained above control:
  Thymus: 99.8%
  
  Results - Strain Distribution:
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Strains Tested: C57BL/6, C3H/He, CBA/J, BALB/c, ATL, AKR/J
  Positive: C57BL/6, C3H/He, CBA/J, BALB/c, ATL
  Negative: AKR