Cell-based Analysis

Cell-Based Assay Kits and Signalling Pathway Detection

Cell-based assay kits and signalling pathway detection reagents for functional cell readouts

You can see whether cells are dividing, dying, differentiating, moving, or responding to a compound or another stimulus. These assays give you a functional readout for a target hypothesis, when profiling compound cytotoxicity, or when a treatment might change a cellular phenotype. This includes viability and cytotoxicity, proliferation and senescence, apoptosis, migration and invasion, adhesion, signalling pathway activity, and phagocytosis.

Match the readout to your endpoint.

1) Cell proliferation, viability, cytotoxicity and senescence. These assays suit a quantitative readout, whether that is cell number, metabolic activity, membrane integrity, or long-term growth behaviour after your treatment.

2) Apoptosis. Apoptosis assays tell regulated cell death apart from other types of toxicity. Early and late markers help you define the mechanism and timing of cell death. No single readout confirms cell death on its own, so combining caspase, MTT, and LDH data distinguishes apoptosis, cytostatic effects, and lytic or necrotic death in your samples.

3) Cell adhesion, migration and invasion. These assays characterise how cells attach, spread, migrate, or invade through two-dimensional or three-dimensional matrices, useful when you need to compare conditions directly.

4) Cell signalling. Signalling assays study pathway activation, receptor engagement, or downstream effector responses, linking these events to functional changes at the cellular level relevant to your model.

5) Phagocytosis. Phagocytosis assays measure how phagocytic cells take up and process particles or other cells, linking directly to immune function and clearance in your system.

Compound cytotoxicity and viability screening

MTT, crystal violet and LDH release assays give viability readouts, pairing well with metabolism assay kits for your dose-response studies.

Apoptosis pathway analysis

Caspase activity marks regulated apoptotic death, while LDH release marks lytic or necrotic death, helping map how your cells die after treatment.

Cell migration and invasion studies

Transwell and gap-closure assays measure directional cell movement, pairing well with cell tracking reagents in your metastasis research.

Cell adhesion assays

Adhesion assays measure how cells attach and spread on a coated surface, characterising integrin-mediated attachment and the compounds you use to block or promote it.

Cell signalling and Rho GTPase activity

Effector domain pull-down kits measure active, GTP-bound Rho GTPases such as RhoA and Rac1, revealing cytoskeletal dynamics downstream of signalling in your pathway of interest.

Phagocytosis assays

Phagocytosis assays measure how macrophages or neutrophils take up and process particles or pathogens, linking the readout to fluorescent imaging and immune clearance.

What is the difference between MTT and LDH cytotoxicity assays?

MTT measures mitochondrial metabolic activity as a proxy for viable, metabolically active cells. A drop in MTT signal can reflect cell death, but it can also reflect a cytostatic effect, where cells stop dividing but stay alive. LDH measures enzyme release from cells with a damaged plasma membrane, so it signals lytic or necrotic death rather than a general drop in metabolic activity.

Neither assay alone tells you the mechanism of cell death. Add a caspase activity assay to detect regulated apoptotic death, since apoptotic cells can stay LDH-negative until late in the process. Combining MTT, LDH, and caspase readouts lets you distinguish cytostatic effects, lytic or necrotic death, and apoptosis with confidence.

How do transwell migration and invasion assays differ?

Both use a membrane insert with set pores. In migration assays, the membrane is left alone and cells move through the pores by chemotaxis. In invasion assays, a reconstituted extracellular matrix layer coats the upper membrane surface.

Cells must degrade this layer before they can cross it. This models the extra barrier that tumour cells face when they invade through the basement membrane.

What controls are needed for a gap-closure migration assay?

You need three controls. First, a mitomycin-C-treated or serum-free control confirms that gap closure reflects migration, not proliferation. Second, a vehicle-only control sets the baseline closure rate.

Third, use a known inhibitor of cell motility as a positive inhibition control. This validates assay sensitivity before you test your experimental compounds.

How is active GTPase distinguished from total GTPase in a pull-down assay?

Effector domain pull-downs use recombinant binding domains, such as PAK1-PBD for Rac1 and Cdc42, or Rhotekin-RBD for RhoA, coupled to beads. These domains bind only the GTP-bound active form.

After pull-down and washing, you detect the bound GTPase by immunoblot. You then normalise it to total GTPase from an aliquot of the same input lysate.

Parent category: Cell-based assay kits

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Cell-Based Assay Kits and Signalling Pathway Detection