CD95 (FAS) Mouse Monoclonal Antibody [Clone ID: UB2]

CAT#: AM26668PU-N

CD95 (FAS) mouse monoclonal antibody, clone UB2, Aff - Purified

Product Images

Flow cytometric analysis of human Fas expression on transfectant. Open histogram indicates the reaction of isotypic control to the cells. Shaded histogram indicates the reaction of AM26668PU-N to the cells.

Specifications

Product Data

• Clone Name: UB2
• Applications: IF, IHC
• Recommended Dilution: Immunohistochemistry on frozen and paraffin sections (see Ref. 8, 9).
  Flow cytometry: 10 μg/mL (final concentration), details see protocol below.
• Reactivities: Human
• Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Recombinant human Fas
• Specificity:

  This antibody recognizes the human Fas antigen specifically.

• Formulation: PBS (pH 7.2)
  State: Aff - Purified
  State: Lyophilized Ig fraction
  Stabilizer: 1% sucrose
  Preservative: 0.09% NaN3
• Reconstitution Method:

  Prepare a stock solution by dissolving the lyophilized antibody in 100 μl of distilled water.

• Purification: Ammonium sulfate precipitation and affinity chromatography on protein A agarose
• Storage:

  Prior to reconstitution store at 2-8°C. Following reconstitution store (in aliquots) at -20°C. Avoid repeated freezing and thawing.
  

• Stability: Shelf life: one year from despatch.

• Database Link:
  Entrez Gene 355 Human
• Background: It is now widely accepted that apoptosis plays an important role in the selection of immature thymocytes and Ag-primed peripheral T cells. Fas antigen is a cell-surface protein that mediates apoptosis. It is expressed in various tissues including the thymus and has structural homology with a number of cell-surface receptors, including tumor necrosis factor receptor and nerve growth factor receptor.
• Synonyms: FASLG receptor, Apo-1 antigen, APT1, FAS1, TNFRSF6
• Note: This product was originally produced by MBL International.
  
  Protocol:

  Flow cytometric analysis for floating cells
  We usually use Fisher tubes or equivalents as reaction tubes for all steps described below.
  1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3].
  2) Resuspend the cells with washing buffer (5 x 106 cells/mL).
  3) Add 50 µL of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
  4) Add 20 µL of Clear Back (human Fc receptor blocking reagent) to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature.
  5) Add 40 µL of the primary antibody at the concentration as suggest in the APPLICATIONS diluted in the washing buffer. Mix well and incubate for 30 minutes at room temperature.
  6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
  7) Add 30 µL of 1:100 FITC conjugated anti-mouse IgG diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature.
  8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
  9) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer.
  (Positive control for Flow cytometry; transfectant)