Cd8b1 Rat Monoclonal Antibody [Clone ID: CT-CD8b]

CAT#: CL012P

Cd8b1 rat monoclonal antibody, clone CT-CD8b, Purified

Product Images

Cell Source: Balb/c thymus labelled with including isotypic control. Percentage of cells stained above control: 90.9%

Specifications

Product Data

• Clone Name: CT-CD8b
• Applications: FC
• Recommended Dilution: Flow Cytometry Analysis (See Protocols).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Mouse CD8 beta.
• Specificity: This anti-Mouse CD8 beta-Chain monoclonal antibody is specific for most thymocytes, cytotoxic/suppressor T-cells and their precursors. The CD8 beta-Chain is also named Ly-3.
• Formulation: PBS
  State: Purified
  State: Liquid purified IgG fraction
  Preservative: 0.09% Sodium Azide
• Concentration: lot specific
• Purification: Affinity Chromatography on Protein G
• Conjugation: Unconjugated
• Storage: Store 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: CD8 antigen, beta chain 1
• Database Link:
  Entrez Gene 12526 Mouse
  UniProt ID P10300
• Background: The CD8 antigen is a cell surface glycoprotein found on most cytotoxic T lymphocytes that mediates efficient cell to cell interactions within the immune system. The CD8 antigen, acting as a coreceptor, and the T cell receptor on the T lymphocyte recognize antigen displayed by an antigen presenting cell (APC) in the context of class I MHC molecules. The functional coreceptor is either a homodimer composed of two alpha chains, or a heterodimer composed of one alpha and one beta chain. Both alpha and beta chains share significant homology to immunoglobulin variable light chains.
• Synonyms: CD8B, CD8B1
• Note: Test Results:
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: BALB/c
  Cell Concentration : 1x10⁶ cells per tests
  Antibody Concentration Used: 2.0 μg/10⁶ cells
  Isotypic Control: Purified Rat IgG2a.
  
  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 1x10e6 cells, representing 1 test).
  4. To each tube, add ~1.0 µg* of this Ab.
  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 (FITC Goat anti-rat IgG (H+L)) at 1:500 dilution.
  9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive).
  10. Wash 2 times at 4°C in media B.
  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).