Flow Cytometry Reagents for Cell Health

The iQue^® Cell Cycle Kit utilizes a DNA binding dye to enable the creation of a cell cycle profile and the quantification of the relative percentage of cells in each phase of the cell cycle.
 

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Key Benefits

• Uses a DNA-specific dye so there is no need to use RNase.
• The dye crosses membranes so there is no need to permeabilize or fix the cells. This also means you can use live cells.
• No wash needed, simply add and read for fast, easily automated workflow.
• Multiplex with other iQue^® kits like iQue^® Cell Membrane Integrity for richer content.

Principle

Cells stained with the cell cycle dye will fluoresce in proportion to their DNA content.

iQue^® Cell Cycle Kit Ordering information

The iQue^® Human 4-Plex Apoptosis Kit is a multi-parameter assay consisting 4 different apoptosis measurements per well (cell viability, caspase 3/7 activity, annexin V binding, and mitochondrial depolarization) for a robust determination of apoptosis progression and insight into mechanism of action. The individual reagent components are also available separately for greater flexibility.

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Key Benefits

• No wash protocol – just add and read for fast, easily automated workflow. Our iQue^® Human 4-plex kit is easier to run than some single endpoint kits!
• Four parameters to deliver rich content, including insight on mechanism of action.
• The 4 components are available separately so you can pick and choose.
• Multiplex with iQue Qbeads^® PlexScreen kits for richer content.

Principle

The activation of Caspase 3 and 7 is detected by the use of NucView 488 Caspase-3/7 substrate, which upon cleavage by activated enzyme, results in an intracellular fluorescent signal.

Surface expression of phosphatidylserine is detected by the binding of Annexin-V to the cell surface.

Cell viability is determined by the ability of live cells to exclude a DNA binding.

Mitochondrial membrane potential is determined by sequestration of a small fluorescent molecule inside the lumen of intact mitochondria with an active membrane potential. Upon mitochondrial depolarization the dye leaks into the cytoplasm and the cell exhibits a decrease in fluorescence.

iQue^® Human 4-Plex Apoptosis Kit Ordering information

iQue^® Human Caspase 3/7 Detection Kit consists of a dye that fluoresces when cleaved by activated caspases 3 & 7 which play an important role in the apoptotic pathway. An increase in fluorescence scores as a positive for apoptotic response. This kit can be multiplexed with other iQue^® kits.

iQue^® Human Annexin V Kit detects phosphatidyl serine on the exterior cell membrane. An increase in fluorescence scores as a positive for apoptotic response. This kit can be multiplexed with other iQue^® kits.

iQue^® Cell Viability Detection Kit consists of a dye that enables to differentiate live cells from dead cells. An increase in fluorescence scores as a positive for apoptotic response. This kit can be multiplexed with other iQue^® kits.

iQue^® Mitochondrial Depolarization Kit consists of a dye that detects mitochondrial depolarization by leaking from the lumen of mitochondria to the cytoplasm. This causes the cell to exhibit decreased fluorescence. This kit can be multiplexed with other iQue^® kits.

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iQue^® Human 4-Plex Apoptosis Kit Ordering information

Each iQue^® Cell Membrane Integrity Dyes consists of a spectrally distinct proprietary dye that is taken up by cells with compromised membranes and excluded by healthy cells with intact membranes. The iQue^® Cell Membrane Integrity Dyes can be multiplexed with other iQue^® reagents.

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Key Benefits

• Different dye options give you more ways to measure cell death.
• No wash protocol, just add and read for fast, easily automated workflow.
• Multiplex with other iQue^® kits like iQue^® Cell Cycle for richer content.

Principle

The iQue^® Cell Membrane Integrity Dyes are fluorescent molecules that are cell impermeant. Healthy cells with intact cell membranes will exclude the dyes. Unhealthy cells with compromised membranes will allow entry of the dye into the intracellular space. Upon entry to the cell, the dye localizes to the nucleus and binds to DNA by intercalation.

iQue^® Cell Membrane Integrity Dyes Ordering information

The iQue^® Cell Encoding Dyes enable labeling of many standard cell lines by using different intensities of a single fluorophore. Each kit includes one dye on a single fluorescent channel that are titrated to three different concentrations to label cells at none, dim, mid, and bright intensity.

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Key Benefits

• Two dye options give you more ways to encode cells.
• No wash protocol, just add and read for fast, easily automated workflow.
• Multiplex with other iQue^® kits like iQue^® Cell Membrane Integrity for richer content.
• Dyes are safe and non-toxic to you and the cells

Principle

Fluorescent cell encoding is a technique where different cell populations are labeled with different intensities of a fluorophore, establishing a unique signature of fluorescence intensity that enables the ability to analyze multiple cell lines simultaneously.

iQue^® Cell Encoding Dyes Ordering information

Each iQue^® Cell Proliferation Dye consists of a spectrally distinct proprietary fluorescent dye that is cell permeable. The iQue^® Cell Proliferation Dyes can be multiplexed with other iQue^® reagents.

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Key Benefits

• Two dye options gives you more ways to measure cell proliferation.
• No wash protocol, just add and read for fast, easily automated workflow.
• Multiplex with other iQue^® kits like iQue Qbeads^® PlexScreen kits for richer content.
• Dyes are safe and non-toxic to you and the cells

Principle

The iQue^® Cell Proliferation Dyes function on the principle of dye dilution. Cell proliferation is detected and quantified based on the halving or “dilution” of the loaded dye after each round of cellular division.

iQue^® Cell Proliferation Dyes Ordering information

iQue^® Cell Proliferation and Encoding (V/Blue) (Tag-it VioletTM) Dye provides a straightforward workflow to simultaneously measures cell proliferation or multiple encoded cell populations. Enable labeling of many standard cell lines by using different intensities of a single fluorophore. Proliferated cells are distinguished from non-proliferated cells by decreased fluorescence intensity.

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Key Benefits

• Ability to encode cell and measure cell proliferation using the violet laser.
• Multiplex with other iQue^® kits like iQue^® Cell Membrane Integrity or iQue Qbeads^® for richer content.
• Dyes are safe and non-toxic to the cells.
• Covalently attaches to intracellular proteins enabling its long-term retention.

Principle

Proliferation Use: The principle of dye dilution. Cell proliferation is detected and quantified based on the halving or “dilution” of the loaded dye after each round of cellular division. Through subsequent cell divisions, daughter cells retain approximately half the fluorescent label of the parent cells.

Encoding Use: Multiple cell populations are stained with different intensities of dye, distinguish up to 4 cell populations (Bright, Mid, Dim and Unstained). Target cells are stained with different concentrations of V/Blue encoding dye in order to discriminate the different cell populations.

iQue^® Cell Proliferation and Encoding (V/Blue) Dye Ordering information

The iQue^® Cell Count and Viability Kit is designed for reproducible quantitative analysis of cell count and viability to screen and profile cells. This assay is optimized to run on the iQue^® platform with BR and VBR configurations combining high throughput sampling and flow cytometry detection capabilities. It provides fast analysis of absolute cell count and viability across a large linear range from a variety of suspension cell lines with a streamlined workflow from cell labeling to analysis.

The kit includes validated reagents and pre-set templates for gating strategy and analysis. Using customer- provided test cells, the kit identifies live cells and determines sample density in a no-wash assay.

This kit is designed for research purposes only.

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Key Benefits

• Increase accuracy and reproducibility — Ensure experimental reproducibility and accuracy to measure viability and absolute cell count.
• Perform high-throughput analysis — Quantify cell viability and count in 96 or 384-well plates across a large linear range without the need for sample dilution.
• Save precious samples — Small sample size (10 µL) assures that precious samples are conserved for more critical downstream assays.
• Maximize productivity — Streamlined workflow with rapid, simple protocol, no washing, no cell dilution.

Workflow

iQue^® Cell Count and Viability Kit Ordering information

The iQue^® Multiplex Antibody Internalization Kits are high throughput, multiplexed, no-wash assays that measures antibody internalization, cell specificity, and cell health using 10 µL sample of cells and 10 µL antibody. Available in 1 x 96, 5 x 96, 1 x 384 and 5 x 384-well format.

The iQue^® Antibody Internalization Reagents are designed to measure internalization of a large number of antibodies in 96- and 384-well formats using 20 µL of cells and 20 µL of labeled antibody.

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Key Benefits

• No wash protocol- fast, easy antibody-labeling method optimal for screening antibody internalization.
• High throughput evaluation of antibody internalization- Assay designed to measure internalization of a large number of antibodies in 96- and 384-well plate formats using 10 µL of cells and 10 µL of labeled antibody (kit).
• Simultaneously measure antibody internalization, cell specificity, and cell health within the same well.
• Available in Human and Mouse species.

Principle

Antibodies are easily labeled with iQue^® Antibody Internalization Reagent, a novel pH sensitive dye. The labeled antibodies have little fluorescence at neutral pH, but become highly fluorogenic at low pH when they are internalized and processed through the acidic lysosome/endosome pathway.

Cell viability can also be measured to assess general cell health and antibody function. Using the same sample, cell specificity can be characterized using encoding dyes (cell populations/types). Data acquisition is performed using the iQue^® 3 and iQue^® Forecyt^® software for data analysis and visualization.

iQue^® Antibody internalization Kits and Reagents Ordering information