CD45 / LCA (CD45RC) Rat Monoclonal Antibody [Clone ID: IBL-8]
Specifications
Product Data | |
Clone Name | IBL-8 |
Applications | FC |
Recommended Dilution | Immunohistochemistry on acetone-fixed frozen sections. Flow Cytometry. Immunoprecipitation. |
Reactivities | Mouse |
Host | Rat |
Isotype | IgG1 |
Clonality | Monoclonal |
Immunogen | Mouse Spleen Cells Donor: Wistar spleen Fusion Partner: Sp-2/0 Ag14 |
Formulation | PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: FITC State: Liquid purified Ig fraction Label: Fluorescein isothiocyanate isomer 1 |
Concentration | 0,1 mg/ml |
Conjugation | FITC |
Background | CD45 (L-CA) is a transmembrane phosphotyrosine phosphatase expressed on leukocytes. Anti-mouse CD45RC is against the exon C-dependent RC isoform and reacts strongly with B cells, and less intensely with most CD8+ T cells. It does not recognize CD4+ T cells. Also, myeloid cells do not express the RC isoform. |
Synonyms | PTPRC, Leukocyte common antigen, L-CA, T200 |
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 1 x 10e6 cells, representing 1 test). 4. To each tube, add 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. (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 by Flow Cytometry Analysis: Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.5 μg/10e6 cells Isotypic Control: FITC Rat IgG1 |
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