Cd4 Rat Monoclonal Antibody [Clone ID: CT-CD4]

CAT#: CL004P

Cd4 rat monoclonal antibody, clone CT-CD4, Purified

Specifications

Product Data

• Clone Name: CT-CD4
• Applications: FC
• Recommended Dilution: Flow Cytometry (See Protocols).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: This antibody recognizes Mouse CD4.
• Formulation: PBS
  State: Purified
  State: Liquid purified IgG fraction
  Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Database Link:
  Entrez Gene 12504 Mouse
• Background: The CD4 (L3/T4) antigen appears to be expressed by the helper/inducer subset of murine T cells and by delayed hypersensitivity T cells but not by cytotoxic T cells or their precursors. CD4 (L3/T4) and CD8a (Ly 2) have been shown to be present on mutually exclusive T cells in the peripheral lymphoid organs but the thymus contains cells expressing both CD4 (L3/T4) and CD8a (Ly 2).
• Synonyms: T-cell surface antigen T4/Leu-3
• 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 2x10⁷ cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1x10⁶ cells, representing 1 test).
  4. To each tube, add ~1.0 µg* of this Ab.
  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 the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C in media B.
  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).