CD71 / TFRC Mouse Monoclonal Antibody [Clone ID: OX-26]

CAT#: SM289B

CD71 / TFRC mouse monoclonal antibody, clone OX-26, Biotin

Product Images

Cell Source: T Cell Blasts - Percentage of cells stained above control:86.2%

Specifications

Product Data

• Clone Name: OX-26
• Applications: FC, IHC
• Recommended Dilution: Flow cytometry.
  Immunohistochemistry on frozen sections.
  Immunoprecipitation.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: PHA activated lymph node cells of PVG rat.
  Donor: BALB/c spleen.
  Fusion Partner: NS1/1.Ag.4.1.
• Specificity: This antibody recognizes the transferrin receptor. This 95kDa molecule is found on proliferating cells and brain endothelium.
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid purified Ig
• Concentration: lot specific
• Purification: Protein G Chromatography
• 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.
• Database Link:
  UniProt ID Q99376
• Background: The transferrin receptor has been structurally characterized as a sulfide bound dimer of identical glycoprotein subunits of 95 kDa. The transferrin receptor is not present on resting blood lymphocytes. On PBL, the receptor appears after activation. The expression of transferrin receptor is coordinately regulated with cell growth. Present on T and B cell lines. The soluble (or serum) transferrin receptor (sTfR) is a circulating truncated form of the membrane receptor protein; it is an 85 kDa glycoprotein forming in serum a 320 kDa complex with diferric transferrin. The most important clinical use of the sTfR determination is in the differential diagnosis between iron deficiency anaemia and the anaemia of chronic disease.
• 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®-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 Ab 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:
  Rat Strain: Wistar
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used:1.0 µg/10e6 cells
  Isotypic Control: Biotin Mouse IgG2a
  
  Cell Source Percentage of cells stained above control:
  Bone Marrow: 14.4%
  T Cell Blasts: 86.2%
  Spleen: 11.6%