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

CAT#: CL039BX

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

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Specifications

Product Data

• Clone Name: 5a-8
• Applications: FC, IHC
• Recommended Dilution: Flow Cytometry (protocol see below).
  Appropriate control samples should always be included in any labelling studies.
• Reactivities: Mouse
• Host: Mouse
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: CBA/J
  Donor: AKR/J Spleen
• Specificity: This antibody detects CD90 (Thy 1.2).
  It reacts with all T lymphocytes from mouse strains expressing the Thy 1.2 phenotype (i.e. C57BL/6, C3H/He, DBA/2, CBA/J, BALB/c), but does not react with lymphocytes expressing the Thy 1.1 phenotype (i.e. AKR/J, B6.PL (74 NS).
• Formulation: PBS containing 0.02% Sodium Azide 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: 0.1 mg/ml
• Purification: Protein G Chromatography.
• Conjugation: Biotin
• 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.
  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 1 x 10e6 cells, representing 1 test).
  4. To each tube, add 0.2-0.5 µg of this antibody 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 (Streptavidin-FITC) at a 1/500 dilution.
  9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C.
  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).
  
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: CBA/J
  Cell Concentration : 1x10e6 cells per tests
  Antibody Concentration Used: 0.2 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgG2b,k
  Cell Source: Percentage of cells stained above control:
  Thymus: 97.8%
  Spleen: 35.4%