Ly6g Rat Monoclonal Antibody [Clone ID: RB6-8C5]

CAT#: CL046B

Ly6g rat monoclonal antibody, clone RB6-8C5, Biotin

Product Images

Cell Source: Bone Marrow Monocytes. Percentage of cells stained above control: 95.3%

Specifications

Product Data

• Clone Name: RB6-8C5
• Applications: FC, IHC, WB
• Recommended Dilution: Flow Cytometry (Ref.1-3).
  Immunohistochemistry on Frozen and Paraffin Embedded Sections (Ref.6).
  Western blot (Ref.5).
  Complement-mediated depletion.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Immunogen: Mouse Granulocytes.
• Specificity: Anti-Mouse Gr-1 monoclonal antibody reacts with the myeloid differentiation antigen Gr-1. (Ref.1,2).
  This 25-30 kDa cell surface antigen is expressed on myeloid cells but not lymphoid or erythroid cells. The expression of the Gr-1 antigen increases with granulocyte maturation (Ref.3) as shown by the distinct populations of bone-marrow cells this monoclonal antibody labels: negative, low positive and high positive. Expression is transient on cells of monocytic lineage (Ref.3).
• Formulation: PBS containing 0.02% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: Biotin
  State: Liquid Ig fraction
• Concentration: 0.1 mg/ml
• Purification: Protein G Chromatography
• Conjugation: Biotin
• Gene Name: lymphocyte antigen 6 complex, locus G
• Database Link:
  Entrez Gene 546644 Mouse
• Synonyms: Gr-1 Granulocyte marker
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with 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 1 x 10⁶ cells, representing 1 test).
  4. To each tube, add 0.1-0.2 µg* of CL046B per 10⁶ 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 FITC Streptavidin at 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:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10⁶ cells per test
  Antibody Concentration Used: 0.1 µg/10⁶ cells
  Isotypic Control: Rat IgG2b
  
  Cell: Source Percentage of cells stained above control:
  Thymus: 1.5%
  Whole Blood: Granulocytes: 90.8%, Monocytes: 91.8%
  Bone Marrow: Granulocytes: 94.2%, Monocytes: 95.3%
  
  Cell Source: Bone Marrow Monocytes
  Percentage of cells stained above control: 95.3%
  
  Strain Distribution by Flow Cytometry Analysis:
  Cell Concentration: 1x10⁶ cells per test.
  Antibody Concentration Used: 0.1 µg/10⁶ cells.
  Strains Tested: BALB/c, C57BL/6, CBA, C3H/He.
  Positive: BALB/c, C57BL/6, CBA, C3H/He.
  Negative: none.