The cytokine human interleukin-2 (IL-2) is a 15kD protein expressed upon activation by peripheral TH1 cells, medullary thymocytes, and a subset of large granular lymphocytes. Secreted IL-2 stimulates proliferation and lymphokine production by T cells, B cells and NK cells.
Suitable for use in Flow Cytometry. Other applications not tested.
Flow Cytometry: 1:50. 80ul labels 5x10e5 cells
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Ficoll prepared human peripheral blood lymphocytes were stimulated 4 hours in the presence of 50ng/ml PMA, 2uM ionomicin and 5uM monensin.
2. Cells were then harvested, washed and fixed for 30 min with 2% buffered formaldehyde after which they were washed two times.
3. Fixed cells were permeabolized 10 minutes in a buffer containing 0.3% Saponin. Subsequent reagent incubations and washes were done using this buffer.
4. 5x10e5 cells per tube were preincubated 5 minutes with 20ul of 0.25mg/ml human IgG (To block nonspecific binding), after which they were incubated 45 minutes on ice with 80ul of I7663-29D at a 1:50 dilution (5ug/ml).
5. Cells were washed three times, fixed and analyzed by FACS. 6. A net 26% sub population of the cells stained positive with a mean shift of 0.94 log10 fluorescent units when compared to a Mouse IgG2a/R-PE negative control (I1904-84B2) at a similar concentration.
7. Binding was blocked when pre incubated 10 minutes with 20ul of 0.5mg/ml anti-IL-2 antibody
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C as an undiluted liquid. Dilute only prior to immediate use. Stable for 12 months at 4 degrees C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.