Ptprc Mouse Monoclonal Antibody [Clone ID: OX-30]

CAT#: CL111R

Ptprc mouse monoclonal antibody, clone OX-30, PE

Product Images

Figure 1. Results-Tissue Distribution.

Specifications

Product Data

• Clone Name: OX-30
• Applications: FC
• Recommended Dilution: Suitable for use in Flow cytometry (See Protocol).
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Lymph node glycoproteins and cells.
• Specificity: This antibody recognizes a monomorphic determinant of the rat leukocyte common antigen (CD45) (1). The antigen recognized is a heavily glycosylated membrane glycoprotein of molecular weight 170,000 kDa on thymocytes but molecular weight 170,000-220,000 kDa on other leukocytes.
• Formulation: PBS with 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: PE
  State: Liquid purified IgG fraction.
  Label: Conjugated to
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography.
• Conjugation: PE
• Database Link:
  Entrez Gene 24699 Rat
• Background: The leukocyte common antigen (L-CA) is a major glycoprotein of haematopoietic cells but is not found on other tissues or erythroid cells. It is present on greater than 95% of thymocytes, bone marrow cells and thoracic duct lymphocytes. This molecule carries much of the carbohydrate of thymocytes and shows interesting heterogeneity amongst T lymphocytes and B lymphocytes. (2,3).
• 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 Rat 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, 0.5 µg-1.0 µg of CL111R or CL111RX 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: (Figure 1)
  Rat Strain: Wistar
  Cell Concentration: 1x10e6 cells per test.
  Antibody Concentration Used: 0.5 µg/10e6 cells.
  Isotypic Control: PE Mouse IgG2a.
  
  Cell-Source Percentage of cells stained above control:
  Thymus: 99.8%
  Spleen: 97.1%
  Lymph Node: 98.9%
  
  Strain Distribution:
  Strains Tested: Wistar, Buffalo, Brown Norway, Fischer 344
  Positive: Wistar, Buffalo, Brown Norway, Fischer 344
  Negative: none