Recognizes the receptor tyrosine kinase, c-kit (CD117). c-kit positive cells are a subset of CD34+ hematopoietic precursor cells and it is expressed on 5-10 % of total adult bone marrow cells.
Flow Cytometry. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with 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 thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 1 µg* of ABIN114257 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 arelight 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 propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media:
Restrictions
For Research Use only
Concentration
0.1 mg/mL
Buffer
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.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing. This product is photosensitive and should be protected from light
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
CD117 or c-kit is a receptor tyrosine kinase. The ligand for this receptor is steel factor (stem cell factor), which exists in both soluble and membrane form. The interaction between steel factor and c-kit is essential for the development of hematopoietic, gonadal and pigment stem cells.Synonyms: KIT, Mast/stem cell growth factor receptor, Proto-Oncogene Tyrosine-Protein Kinase, SCFR