Cd28 Mouse Monoclonal Antibody [Clone ID: JJ319]

CAT#: SM268PX

Cd28 mouse monoclonal antibody, clone JJ319, Purified

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Specifications

Product Data

• Clone Name: JJ319
• Applications: FC, IP
• Recommended Dilution: Flow cytometry.
  Immunoprecipitation.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat CD28 transfected A20J
  Donor: BALB/c spleen
  Fusion Partner: X63-Ag8.653
• Specificity: Anti-rat CD28 monoclonal antibody recognizes a 90kDa homeodimeric cell surface glycoprotein. CD28 has been found to be a potent costimulatory receptor on T cells. It is expressed on all peripheral rat ab and most gd T cells, as well as on approximately half of all NK cells.
  This clone can costimulate T cell proliferation and IL-2 secretion by resting rat T cells.
• Formulation: PBS containing 0.02% sodium azide (NaN3) as preservative
  State: Purified
  State: Liquid purified Ig fraction
• Concentration: 1,0 mg/ml
• Purification: Affinity chromatography on Protein G
• Gene Name: Rattus norvegicus Cd28 molecule (Cd28)
• Database Link:
  Entrez Gene 25660 Rat
• Synonyms: TP44
• 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®-Rat 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 1x10e6 cells, representing 1 test).
  4. To each tube, add 1.0 μg* of this antibody.
  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 (PE Goat anti-mouse IgG
  (H+L)) at 1/50 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).
  Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Rat Strain: Fischer
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 μg/10e6 cells
  Isotypic Control: Mouse IgG1
  Percentage of cells stained above control:
  Thymus 18.0%
  Splenic T Cells* 71.7%
  *(T cells isolated with Rat T Cell Recovery Column Kit)