CD45 / LCA Rat Monoclonal Antibody [Clone ID: IBL-5/25]

CAT#: AM31886PU-N

CD45 / LCA rat monoclonal antibody, clone IBL-5/25, Purified

Product Images

Cell Source: Lymph Node Percentage of cells stained above control: >95% (Representative Histogram)

Specifications

Product Data

• Clone Name: IBL-5/25
• Applications: FC, IHC, IP
• Recommended Dilution: Immunohistochemistry on aceton-fixed frozen sections.
  Immunoprecipitation.
  Flow Cytometry.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: IL-3 dependent mast cells derived from WB- +/+ mice
  Donor: Wistar spleen
  Fusion Partner: X63.653. Ag8
• Specificity: Anti-mouse CD45 monoclonal antibody detects CD45 (L-CA) which is a transmembrane phosphotyrosine phosphatase expressed on leukocytes. This mAb induces the in vitro clustering of mouse lymphocytes (both T and B cells).
• Formulation: PBS containing 0.02% sodium azide (NaN3) as preservative
  State: Purified
  State: Liquid purified Ig fraction
  Label: Fluorescein isothiocyanate isomer 1 (FITC)
• Concentration: 1,0 mg/ml
• Purification: Affinity chromatography on Protein G
• 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 2.0 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 (FITC Goat anti-rat IgG (H+L)) at 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: BALB/c
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 2.0μg/10e6 cells
  Isotypic Control: Purified Rat IgG1
  Percentage of cells stained above control:
  Mesenteric Lymph Node Cells 95%