HiCK-2 Human Cytokine Positive Control Cells
- Brand BD Pharmingen™
- Concentration 5x10^6 cells/ml
Intracellular staining (flow cytometry) (Routinely Tested)
- Storage Buffer Frozen in FBS and 10% DMSO.
- Regulatory Status RUO
Regulatory Status Legend
This suspension contains Human intracellular CytoKine-2 (HiCK-2) Positive Control Cells. The HiCK-2 frozen cell suspension contains fixed, non-permeabilized human lymphoid cells. The suspension includes cells that express detectable levels of intracellular IL-3, IL-4, IL-10, IL-13 and GM-CSF as determined by immunofluorescent intracellular cytokine staining and flow cytometry. HiCK-2 cell suspensions were prepared by stimulating human PBMCs in the presence of a protein transport inhibitor. After stimulation, the cells were harvested and fixed, then stored in 1 mL of 10% dimethylsulfoxide and 90% fetal bovine serum at -80°C. HiCK-2 cells contain a measurable proportion of cytokines, with representative flow cytometric data shown below. Performance from individual lots of HiCK-2 cells may differ due to donor variability. Investigators should anticipate similar, though not identical, results to those shown below.
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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.
- 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.
- 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/pharmingen/protocols for technical protocols.
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.