BD, 555062, BD Pharmingen™ HiCK-2 Human Cytokine Positive Control Cells

Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 5x10^6 cells/ml
RRID: AB_2869016
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. After thawing and thoroughly resuspending cells with a pipette, “single-use” aliquots can be refrozen at -80°C and stored in polypropylene microtubes for use at a later time.
Recommended Assay Procedures: Recommended Assay Procedures Flow Cytometry: HiCK-2 Cytokine Positive Control Cell suspensions contain intracellular IL-3, IL-4, IL-10, IL-13 and GM-CSF which have accumulated and are detectable by intracellular flow cytometric analysis. These cells may serve as a positive control for verifying anti-cytokine antibody performance and/or the flow cytometric staining procedure itself (e.g. permeabilization).  For flow cytometric 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.  HiCK-2 cells are supplied fixed and non-permeabilized in dimethylsulfoxide (DMSO), so should be washed at least twice with staining buffer to remove the DMSO. The cells must then be permeabilized by incubating for 10-15 min in BD Perm/Wash™ buffer (Cat. No. 554723), spun down to pellet and then followed by at least one wash in BD Perm/Wash™ buffer. Cells can then be stained with either PE Rat Anti-Human IL-3 (Cat. No. 554676), PE Mouse Anti-Human IL-4 (Cat. No. 554516), PE Rat Anti-Human IL-10 (Cat. No. 554706), PE Rat Anti-Human IL-13 (Cat. No. 554571) or PE Rat Anti-Human GM-CSF (Cat. No. 554507).  Investigators should note that for optimal detection of IL-10 or IL-13, ≥ 20,000 events should be acquired on the flow cytometer. Note: Cytokine-specific antibody staining of HiCK-2 cells can be demonstrated by preincubation of conjugated cytokine-specific antibody with recombinant cytokine or by pretreatment of the HiCK-2 cells with unlabeled (purified) blocking antibody.  Investigators should note that variation with cell activation may contribute to suboptimal blocking.
Product Notices: Product Notices This product contains human blood, serum, cells, or materials derived from them, which are potentially hazardous materials. Use universal precautions when handling. Handle as if product were capable of transmitting disease. Material used in this product has been tested using FDA approved methods and found negative for Human Immunodeficiency Virus (HIV-1/HIV-2), Hepatitis B Surface Antigen (HBSAG) and antibody to Hepatitis C Virus (HCV). However, no known test method can offer complete assurance that specimens of human origin will not transmit infectious disease. When handling or disposing, follow precautions described in CDC and FDA recommendations and OSHA Bloodborne Pathogen recommendations. Product Use Limitations: This product is for research use only. Research products are labeled as Research Use Only (RUO) and are not for use in diagnostic or therapeutic procedures. Customer agrees not to use the product for purposes which allow for any identification of the individual donors. Customer agrees to notify BDB Technical Services within five (5) business days of becoming aware of any identification or re-identification of an individual in connection with this product. Avoid contact with skin and eyes. Source of all serum proteins is from USDA inspected abattoirs located in the United States. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.