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

CAT#: CL050FX

Neutrophils rat monoclonal antibody, clone 7/4, FITC

Specifications

Product Data

• Clone Name: 7/4
• Applications: FC
• Recommended Dilution: This antibody is suitable for use in Flow cytometry.
  This clone has also been reported to be useful for Immunohistochemistry (both frozen and paraffin sections).
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Cultured bone marrow cells.
• Specificity: This antibody is specific for detecting mouse neutrophils.
  Strains reported to be positive for the 7/4 clone are: AKR, C57BL/6, C57BL/10,C58, DBA/2, MF1, NZB, NZW, SJL, Swiss (PO) and 129J.
  Strains reported to be negative/weak for the 7/4 clone are: A2G, A/Sn, ASW, BALB/c, C3H/HEH and CBA.T6T6.
• Formulation: PBS with 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: FITC
  State: Liquid Ig fraction.
• Concentration: 0.1 mg/ml
• Conjugation: FITC
• Note: Actual results of Flow cytometric analysis
  Mouse strain: C57BL/6
  Cell source: Peripheral Blood Leukocytes
  Cell concentration: 1x10e6 cells per test
  Antibody concentration: 1.0 µg/10e6 cells
  Isotypic control: FITC Rat IgG2a
  Percentage of cells stained above control: 21.84%
  
  Protocol: FLOW CYTOMETRY ANALYSIS:
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population using a 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 1 x 10e6 cells,
  representing 1 test).
  4. To each tube, add ~1.0 μg of CL050F 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).