Apoptosis Pathway Assays and Profiling Services

CD Biosynsis offers comprehensive Apoptosis Pathway Assays essential for drug screening, mechanism of action studies, and toxicology assessments. Apoptosis, or programmed cell death, is tightly regulated and plays a critical role in development, disease, and response to therapeutic agents. Our platform allows for precise quantification of multiple apoptosis hallmarks, from initial activation (caspase activity, mitochondrial depolarization) to late-stage events (DNA fragmentation, membrane asymmetry). We utilize highly sensitive, high-content analysis (HCA), flow cytometry, and enzyme activity assays to deliver quantitative and cell-specific data. Researchers can select and combine individual assay modules to build a custom profiling panel, enabling rapid and detailed characterization of compounds targeting the intrinsic, extrinsic, or executioner phases of the apoptotic pathway.

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Multiparametric Analysis of Programmed Cell Death

The decision for a cell to undergo apoptosis is governed by complex signaling networks involving the BCL-2 family, adaptor proteins, and the Caspase cascade. Accurate drug profiling requires monitoring multiple, sequential events to distinguish true apoptosis from necrosis or ferroptosis. Our integrated approach enables the simultaneous tracking of early, mid, and late-stage apoptotic markers. By offering selectable modules, researchers gain flexibility to focus on specific phases—for example, measuring the activation of initiator Caspases (8 and 9) or quantifying mitochondrial outer membrane permeabilization (MOMP). This targeted and comprehensive analysis ensures high-quality mechanistic data, minimizing ambiguity in compound efficacy and toxicity testing.

Apoptosis Pathway Assay Workflow

A staged approach for multiparametric detection of programmed cell death.

Cell Culture and Compound Treatment

Early Stage (Initiation) Analysis

Mid Stage (Execution) Analysis

Late Stage and Data Reporting

Cell Preparation: Seed selected cell line (adherent or suspension) into HTS-compatible plates.

Compound Dosing: Treat cells with varying concentrations of test compounds and controls for kinetic time points (e.g., 6, 12, 24 hours).

Mitochondrial Integrity: Assess MMP loss using fluorescent probes (e.g., JC-1, TMRM) via plate reader or HCI (if selected).

BCL-2 Family/ROS: Quantify activation of pro-apoptotic proteins or generation of reactive oxygen species (if selected).

Caspase Activity: Measure enzymatic activity of selected Caspases (e.g., 3/7, 8, 9) using fluorogenic substrates in lysate or live cells.

Protein Cleavage: Confirm Caspase activation by Western Blot or HCI detection of cleaved substrates (e.g., Cleaved Caspase-3, Cleaved PARP).

Membrane Asymmetry: Perform Annexin V/Propidium Iodide (PI) staining for flow cytometry or HCI analysis.
DNA Fragmentation: Execute TUNEL assay for definitive confirmation of late-stage apoptosis (if selected).
Reporting: Deliver dose-response curves, kinetic analysis, and multiparametric plots (e.g., Annexin V vs. PI).

High Content Analysis for Apoptotic Mechanisms

Multiparametric Analysis

Ability to simultaneously monitor 3+ apoptotic events (MMP, Caspase, Annexin V) for clear MoA.

High Content Imaging (HCI)

Single-cell resolution for assessing nuclear morphology and distinguishing apoptosis from necrosis.

Caspase Kinetic Profiling

Quantitative enzyme assays for determining the IC50/Ki of inhibitors against specific Caspase targets.

Customizable Panels

Flexibility to combine early, mid, and late-stage assays to precisely map a compound's pathway activation.

Client Testimonials on Apoptosis Assays

"The combination of Caspase-8 activity and Cytochrome c release assays definitively proved our compound activated the extrinsic pathway, accelerating our MoA studies."

Dr. Lena Olin, Oncology Research Director

"Using the Annexin V/PI flow cytometry service, we clearly distinguished apoptotic cells from necrotic cells, which was vital for accurately determining the therapeutic window."

Mr. Kenji Sato, Toxicology Screening Lead

"The High Content Imaging platform provided single-cell data on nuclear condensation and MMP loss, giving us superior mechanistic insight compared to bulk lysate assays."

Ms. Clara Davis, Translational Biology Manager

FAQs about Apoptosis Pathway Assays

Still have questions?

What is the difference between Annexin V and TUNEL staining?

Annexin V staining (Phosphatidylserine externalization) marks an early to mid-stage event of apoptosis. TUNEL staining (DNA fragmentation) marks a late-stage, irreversible executioner event.

What does Caspase-8 activity indicate?

Caspase-8 is the primary initiator caspase for the extrinsic (death receptor-mediated) pathway of apoptosis. Its activation suggests the compound is acting through cell surface receptors (e.g., Fas, TNF).

Can you distinguish apoptosis from necrosis?

Yes. Apoptosis is characterized by Annexin V positivity and morphological changes (nuclear condensation), while necrosis is characterized by cell swelling and early loss of membrane integrity, which is typically detected using dyes like Propidium Iodide (PI) or Calcein AM/Ethidium Homodimer.

What is mitochondrial outer membrane permeabilization (MOMP)?

MOMP is a critical early event in the intrinsic apoptosis pathway, leading to the release of pro-apoptotic factors like Cytochrome c into the cytosol, which in turn triggers the Caspase cascade via the apoptosome.

How much does Metabolic Engineering services cost?

The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.

Do your engineered strains meet regulatory standards?

We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).

What to look for when selecting the best gene editing service?

We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.

Does gene editing allow customisability?

Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.

What is the process for keeping data private and confidential?

We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.

Apoptosis Pathway Assays and Profiling Services

Multiparametric Analysis of Programmed Cell Death

Selectable Apoptosis Pathway Assay Modules

Early Stage Events and Signaling

Choose Initial Apoptosis Markers for Detection

Caspase Cascade Activation

Choose Specific Caspase Targets for Enzyme Activity

Late Stage and Executioner Events

Select Final Apoptosis Markers for Confirmation

Apoptosis Pathway Assay Workflow

High Content Analysis for Apoptotic Mechanisms

Client Testimonials on Apoptosis Assays

FAQs about Apoptosis Pathway Assays