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

CAT#: AM26037BT-N

Il2ra rat monoclonal antibody, clone PC61.5.3, Biotin

Product Images

Cell Source: T Cell Blasts (Con A Activated) Percentage of cells stained above control: 99.4%

Specifications

Product Data

• Clone Name: PC61.5.3
• Applications: FC, IHC, IP
• Recommended Dilution: Immunoprecipitation.
  Immunohistochemistry on frozen sections.
  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: Anti-mouse CD25 (IL-2R) monoclonal antibody reacts with the low affinity alpha chain of the interleukin-2 receptor antigen present on activated T and B cells in mice. CL8925B 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: Biotin
  State: Liquid purified Ig fraction
• Concentration: lot specific
• Purification: Affinity chromatography on Protein G
• Conjugation: Biotin
• Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Gene Name: interleukin 2 receptor, alpha chain
• Database Link:
  Entrez Gene 16184 Mouse
  UniProt ID P01590
• 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 0.2-0.1 μg* of AM26037BT-N or AM26037BT-L 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 (Streptavidin-FITC) at a
  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 tests
  Antibody Concentration Used: 0.1 μg/10e6 cells
  Isotypic Control: Biotin Rat IgG1
  Cell Source Percentage of cells stained above control:
  Cell Blasts (Con A activated) 99.4%
  Thymus (unactivated) 1.7%