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

CAT#: CL046FX

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

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Specifications

Product Data

• Clone Name: RB6-8C5
• Applications: FC, WB
• Recommended Dilution: Flow cytometry (1,2,3).
  Western blot (5).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2b
• Clonality: Monoclonal
• Specificity: This antibody reacts with the Mouse myeloid differentiation antigen GR-1 (1,2).
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: FITC
  State: Liquid Ig fraction
  Absorption emission: 495 nm / 528 nm
• Concentration: 0.1 mg/ml
• Purification: Protein G chromatography
• Conjugation: FITC
• Gene Name: lymphocyte antigen 6 complex, locus G
• Database Link:
  Entrez Gene 546644 Mouse
• Background: GR-1 is a 25-30 kDa cell surface antigen and is expressed on myeloid cells but not lymphoid or erythroid cells. The expression of the Gr-1 antigen increases with granulocyte maturation (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 (3).
• 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.
  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 0.1 - 0.5 µg of antibody 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. (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 by Flow Cytometry Analysis:
  Mouse Strain: CBA/J
  Cell Concentration : 1x10e6 cells per test
  Antibody Concentration Used: 0.1 µg/10e6 cells
  Isotypic Control: FITC Rat IgG2b
  Cell Source Percentage of cells stained above control:
  Thymus 1.5%
  Whole Blood Monocytes 87.2%
  Bone Marrow Macrophages 90.0%
  
  (see picture below)
  
  Strain Distribution by Flow Cytometry Analysis:
  Procedure: see above
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.1 µg/10e6 cells
  Strains Tested: BALB/c, C57BL/6, CBA, C3H/he, AKR
  Positive: BALB/c, C57BL/6, CBA, C3H/he, AKR
  Negative: none