Il2ra Rat Monoclonal Antibody [Clone ID: PC61.5.3]

CAT#: AM26037FC-S

Il2ra rat monoclonal antibody, clone PC61.5.3, FITC

Product Images

Cell Source: T Cell Blasts (Con A Activated) Percentage of cells stained above control: 90.9%

Specifications

Product Data

• Clone Name: PC61.5.3
• Applications: FC
• Recommended Dilution: Immunoprecipitation.
  Flow Cytometry.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: B6.1 CTL Cell line
  Donor: OFA rat spleen
  Fusion Partner: mouse myeloma line P3X63Ag8.653
• Specificity: This monoclonal antibody reacts with the low affinity alpha chain of the Interleukin-2 receptor antigen present on activated T and B cells in mice.
  AM26037FC inhibits IL-2 binding and IL-2 dependent proliferation.
• Formulation: PBS containing 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: FITC
  State: Liquid purified Ig fraction
  Label: Fluorescein isothiocyanate isomer 1
• Concentration: 0.1 mg/ml
• Purification: Affinity Chromatography on Protein G
• Conjugation: FITC
• Gene Name: Mus musculus interleukin 2 receptor, alpha chain (Il2ra)
• Database Link:
  Entrez Gene 16184 Mouse
• Synonyms: Interleukin-2 receptor alpha chain, IL-2 receptor alpha subunit, IL-2-RA, IL2-RA, p55, TAC antigen
• 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 0.5-1.0 μg of AM26037FC 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 tests
  Antibody Concentration Used: 1.0 μg/10e6 cells
  Isotypic Control: FITC Rat IgG1
  Cell Source Percentage of cells stained above control:
  T Cell Blasts (Con A activated) 90.9%
  Thymus (unactivated) 3.8%