Cd28 Mouse Monoclonal Antibody [Clone ID: JJ319]

CAT#: SM268FX

Cd28 mouse monoclonal antibody, clone JJ319, FITC

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Specifications

Product Data

• Clone Name: JJ319
• Applications: FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat CD28 transfected A20J cells
  Donor: BALB/c spleen
  Fusion Partner: X63-Ag 8.653
• 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 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: lot specific
• Purification: Affinity chromatography on Protein G
• Conjugation: FITC
• 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: Cd28 molecule
• Database Link:
  Entrez Gene 25660 Rat
  UniProt ID P31042
• 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 1 x 10e6 cells, representing 1 test).
  4. To each tube, add 1.0 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:
  Rat Strain: Fischer
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 μg/10e6 cells
  Isotypic Control: FITC Mouse IgG1
  Percentage of cells stained above control:
  Thymus 36.3%
  Splenic T Cells* 79.7 %
  *(T cells isolated with Rat T Cell Recovery Column Kit)