BD, 554653, BD Pharmingen™ MiCK-2 Mouse Cytokine Positive Control Cells

Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 5x10^6 cells/ml
RRID: AB_2869003
Storage Buffer: Frozen in FBS and 10% DMSO.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store product at -80°C prior to use or for long term storage of stock solutions. Rapidly thaw and quick-spin product prior to use. Avoid multiple freeze-thaws of product. This preparation contains no preservatives, thus it should be handled under aseptic conditions. Upon receipt, store the cell suspension at -80°C. Alternatively, the frozen cell suspension can be thawed and the capped vial should be quick spun to ensure complete retrieval of the contents from the vial. After thoroughly resuspending cells with a pipette, "single-use" aliquots can be refrozen and stored (-80°C) in polypropylene microtubes for a use at a later time.
Recommended Assay Procedures: Recommended Assay Procedures MiCK-2 Positive Control Cell suspensions are intended to provide cells that contain intracellular accumulations of IL-4, IL-10 or GM-CSF which are easily detectable by immunofluorescent staining of intracellular cytokines and flow cytometry. As such, these cells serve as positive controls for verifying the activity of fluorescent anti-cytokine antibodies and the staining procedure itself (eg, permeabilization). For staining, the frozen cell preparation should first be quickly and carefully thawed. Aliquots of the cell suspension can then be transferred to microwells or tubes. The fixed and non-permeabilized cells should be washed twice with staining buffer to remove the DMSO. It is recommended that the washed cells first be stained with a fluorescent conjugate of RM4-5 (eg, Cat. No. 553047, 553049, 553050 or 553051), an antibody which is known to bind to CD4 molecules expressed on the surface of fixed mouse CD4+ T cells. The cells must then be incubated for 10 min in permeabilization/wash buffer and washed. The cells are then stained with a fluorescent conjugate of either BVD4-1D11 (rat anti-mouse IL-4; Cat. No. 553051), 11B11 (rat anti-mouse IL-4; Cat. No 554435 or 554436), JES5-16E3 (rat anti-mouse IL-10; Cat. No. 554466, 554467, or 554468) or MP1-22E9 (rat anti-mouse GM-CSF; Cat. No. 554406). Note: Cytokine-specific antibody staining of MiCK-2 cells can be demonstrated by preincubation of conjugated cytokine-specific antibody with recombinant cytokine or by pretreatment of the MiCK-2 cells with unlabled blocking antibody. Due to the cell activation procedure, a small proportion of cells may stain non-specifically (ie, not blocked with either unlabeled antibody or ligand).