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

CAT#: AM31886FC-N

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

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
• 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 nd 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
• 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 – 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: 2.0μg/10e6 cells
  Isotypic Control: FITC Rat IgG1
  Percentage of cells stained above control:
  Mesenteric Lymph Node Cells 95%