Cell Based Assays

Easy to use, caspase inhibitor-based apoptosis detection kits that have been tested in primary neurons and many other cell types! Features:

•  Easy-Just add the reagent directly to the cell culture and incubate (see sidebar).
•  Fast-The reactions start within 15 minutes of addition to the cells, however we recommend an incubation of 1-4 hours.
•  Accurate-There is no interference from pro-caspases or inactive forms of the enzyme. Quantitative Read with a fluorescence plate reader, microscope, or flow cytometer.
•  Reliable-Only cells with active caspase enzymes fluoresce. The reagents may be incubated up to 72 hours without killing the cells
•  Sensitive-Detects background levels of apoptosis in controls. Detect apoptosis earlier than Annexin V and TUNEL Caspases are released before the phospholipid turnover & DNA laddering.

The chymotrypsin-like serine rotease subfamily plays a key role in a number of diverse physiological processes, including the maintenance of homeostasis, inflammation and immune reactions, apoptosis, and cell survival. For example,chymotrypsin-like serine proteases have been shown to be activated during apoptosis signaling. Additionally, some chymotrypsin serine protease inhibitors have been shown to induce apoptosis by blocking IkBa degradation, which is important in the NFkB cell survival signaling pathway.

Use our FFCK kit to detect chymotrypsin-like serine proteases that favor phenylalanine;use the FLCK kit for those that favor leucine; use the kits with an additional spacer sequence to reduce steric hindrance at the binding site of the serine protease.

Just culture your cells, add MR to the media, and incubate. If the enzymes are active, they will cleave the reagent causing it to fluoresce red. MR kits are true substrate assays – the red fluorescent label is quenched until it is actually cleaved by active enzymes. Only cells with active enzymes will fluoresce, so you don’t get any signal from pro-enzymes or inactive forms of the enzyme.

Cytolytic activity is an important process by which the immune system eliminates compromised cells in the body. Whether describing antibody-dependent cell-mediated cytotoxicity (ADCC), complement-mediated cytotoxicity, or lymphocyte-mediated cytotoxicity, the activation of the immune system leads to the removal of target cells infected by pathogens or transformed by cancer. Alternatively, cytotoxicity can simply refer to any compound (drug or toxin) that also results in killing of the target cells.

We offer two easy, reliable, and accurate assays to measure cytotoxicity. The Basic Cytotoxicity Test simplifies measurement of necrotic and membrane-compromised cells. The Total Cytotoxicity Test  further distinguishes apoptosis from necrosis, leading to a more accurate assessment of cytolytic activity. At least 10% more cytolytic activity is often measured when apoptotic cells are also identified. ICT’s kits provide a direct measurement of cytotoxicity rather than using an indirect indicator, such as the release of LDH or ATP enzymes, and there is no need to lyse the cells. These assays do not use any radioisotopes, they are much safer and more cost effective to run than 51Cr assays. Highlights:

• Easy – run samples in a single tube
• Safe – no radioactivity
• Fast – finish in one day Sensitive – measure killed and apoptotic cells
• Accurate – analyze cells with a flow cytometer

Autophagy is a conserved lysosomal recycling process by which cells break down their own components such as proteins, lipids, and carbohydrates. The process plays an important role in maintaining homeostasis and destroying intracellular pathogens. In addition, autophagy can be upregulated in times of starvation or stress to provide additional nutrients for the cell. Dysregulation of autophagy has implications in cancer, infection, and degenerative diseases.

Autophagy is a three stage process. First, cytoplasmic components targeted for degradation are sequestered, resulting in the formation of the autophagosome. Next, the autophagosome fuses with the lysosome to form the autophagolysosome or autolysosome. Finally, degradation of the autophagosomal contents occurs.

Our Autophagy Assay, Red (cat# KF17373) enables researchers to detect and monitor the in vitro development of autophagy in living cells. The Autophagy Probe is cell-permeant and fluoresces red when inserted in the lipid membranes of autophagosomes and autolysosomes. Results can be read using a flow cytometer.

• They are non-magnetic soluble protein complexes generated by multimerization of αCD3- and αCD28 Fab-Streps with a Strep-Tactin® multimer.
• They are completely reversible reagents, i.e. they can be removed from the cells by the addition of biotin.
• The Strep-Tactin® CD3/CD28 premix for T cell expansion is easy and fast with ready-to-use reagents.
• FACS Data available for the expansion of T-Cells using CD4+, CD8+, CD25 and CD 65 markers.

Fluorescent Fab-TACS®, complexes of Strep-tagged Fab fragments (Fab-Streps) and Strep-Tactin® PE or APC, are directed against cell surface markers including those of T cells (CD3, CD4, CD8), memory T cells (CD8, CD62L, CD45RA-), regulatory T cells (CD4, CD25, CD45RA).

These products can also be used in magnetic T-Cell Isolation.