Thy1 Mouse Monoclonal Antibody [Clone ID: 5a-8]
Specifications
Product Data | |
Clone Name | 5a-8 |
Applications | FC |
Recommended Dilution | Flow Cytometry. |
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, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml Label: PE State: Liquid purified IgG |
Concentration | 0.1 mg/ml |
Purification | Protein G Chromatography |
Conjugation | PE |
Database Link | |
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 0.5 µg* of this Ab. 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: 0.5 µg/10e6 cells Isotypic Control: PE-Mouse IgG2b,k Cell Source Percentage of cells stained above control: Thymus: 98.4% Spleen: 28.8% Results - Strain Distribution: Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.5 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/J |
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