Neutrophils Rat Monoclonal Antibody [Clone ID: 7/4]

CAT#: CL050RX

Neutrophils rat monoclonal antibody, clone 7/4, PE

Product Images

Cell Source: Peripheral Blood Leukocytes - Percentage of cells stained above control: 11.4%

Specifications

Product Data

• Clone Name: 7/4
• Applications: FC
• Recommended Dilution: Flow cytometry (see protocol).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Cultured bone marrow cells
• Specificity: Reacts with mouse neutrophils. Strains to be positive for the 7/4 clone are: AKR, C57BL/6, C57BL/10, C58, DBA/2, MF1, NZB, NZW, SJL, Swiss (PO), 129J.
  Strains reported to be negative/weak for the 7/4 clone: A2G, A/Sn, ASW, BALB/2, C3H/HEH and CBA.T6T6.
• Formulation: PBS with 0.09% NaN3 as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml
  Label: PE
  State: Liquid
  Label: conjugated
  Absorption emission: 488 nm / 575 nm
• Concentration: 0.1 mg/ml
• Purification: Protein A affinity purified Ig
• Conjugation: PE
• Note: This clone has also been reported to work in Western blotting and immunohistochemistry (frozen and paraffin sections, 2).
  
  Protocol: FLOW CYTOMETRY ANALYSIS:
  
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population using an NH4Cl lysing buffer.
  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 CL050R 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:
  Mouse Strain: C57BL/6
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 1.0 µg/10e6 cells
  Isotypic Control: PE Rat IgG2a