CD44 Mouse Monoclonal Antibody [Clone ID: OX-49]

CAT#: CL110F

CD44 mouse monoclonal antibody, clone OX-49, FITC

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Product Data

• Clone Name: OX-49
• Applications: FC, IHC
• Recommended Dilution: Flow cytometry (See protocol).
  Immunohistochemistry on Frozen Sections and Paraffin Sections.
• Reactivities: Rat
• Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: T cell blasts.
• Specificity: This anti-Rat CD44 monoclonal antibody recognizes an epitope on both standard CD44 and its splice variant.
• Formulation: PBS with 0.02% sodium azide as preservative and EIA grade BSA as a stabilizer
  Label: FITC
  State: Liquid purified IgG fraction
• Concentration: 0.1 mg/ml
• Purification: Protein G affinity chromatography
• Conjugation: FITC
• Background: This antigen is expressed on most leukocytes (except a sub population of B cells) and increases upon activation. This antibody binds extracellularly to the standard (S) form on rat leukocytes, but it is not known if they bind to the N-terminal region. It has also been reported that the antibody may bind to melanoma cell lines that express CD44V (splice variant form).
  CD44 is expressed on most leukocytes except a sub population of B cells. Its expression is increased on T and B blasts.
• Synonyms: LHR, MDU2, MDU3, MIC4, CDw44, Epican, ECMR-III, HUTCH-I, Heparan sulfate proteoglycan, Hermes antigen, Hyaluronate receptor, PGP-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 Rat 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 1.0-0.5 µg* of CL110F or CL110FX.
  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:
  Rat Strain: Wistar
  Cell Concentration: 1 x 10e6 cells per tests
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Isotypic Control: FITC Mouse IgG2a.
  
  Cell Source Percentage of cells stained above control:
  Thymus: 99.6%
  Spleen: 75.9%
  Lymph Node: 96.8%
  
  N.B. Appropriate control samples should always be included in any labelling studies.
  * For optimal results in various applications, it is recommended that each investigator determine dilutions appropriate for individual use.