Tfrc Rat Monoclonal Antibody [Clone ID: R17217.1.4]

CAT#: AM00702BT-L

Tfrc rat monoclonal antibody, clone R17217.1.4, Biotin

Specifications

Product Data

• Clone Name: R17217.1.4
• Applications: FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: Anti-mouse CD71 antigen monoclonal antibody reacts with the transferrin receptor (CD71). The expression of CD71 is up-regulated after B cell or T cell activation.
• Formulation: PBS containing 0.09% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein
  Label: Biotin
  State: Liquid purified Ig fraction
• Concentration: lot specific
• 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: transferrin receptor
• Database Link:
  Entrez Gene 22042 Mouse
  UniProt ID Q62351
• Synonyms: TfR1, p90, Transferrin receptor protein 1
• 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 ~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.
  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:
  (Representative Histogram)
  Mouse Strain: Mouse Hybridomas
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 μg/10e6 cells
  Isotypic Control: Biotin Rat IgG2a