In Vitro Efficacy Testing Services for Systemic Mastocytosis

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Systemic Mastocytosis. Our service enables comprehensive evaluation of candidate compounds targeting abnormal mast cell proliferation and activation, which are central to this rare hematologic disorder. Key targets relevant to Systemic Mastocytosis include the KIT receptor tyrosine kinase, associated downstream signaling pathways, and mediators of mast cell degranulation. Our assays are designed to assess pathological processes such as cell proliferation, survival, and mediator release that drive disease progression.

We offer a diverse range of cell-based and biochemical in vitro assays to evaluate compound efficacy, mechanism of action, and cellular responses in Systemic Mastocytosis models. These methods enable high-throughput and quantitative analysis of drug effects on relevant molecular targets and cellular functions.

ATP assay: Measures cellular ATP levels as an indicator of cell viability and proliferation, providing a sensitive readout for cytotoxicity and growth inhibition.

Bioluminescence resonance energy transfer (BRET) assay: Detects protein-protein interactions and signaling events in live cells using bioluminescent donor and fluorescent acceptor molecules.

Chemiluminescent assay: Utilizes chemiluminescent substrates to quantify specific biological activities, such as enzyme function or molecule binding, with high sensitivity.

Fluorescence resonance energy transfer (FRET) assay: Monitors molecular interactions and conformational changes in real time by measuring energy transfer between fluorescent probes.

Fluorescent assay: Employs fluorescent dyes or probes to assess cellular processes including viability, apoptosis, or mediator release.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Combines time-resolved fluorescence detection with antibody-based recognition to quantify biomolecules or protein interactions in a homogeneous format.

Rhodamine 123 accumulation assay: Evaluates mitochondrial function and drug efflux activity by measuring the accumulation of the fluorescent dye rhodamine 123 in cells.

We measure a variety of key pharmacological parameters to characterize compound potency, binding affinity, and antimicrobial activity. These parameters are essential for ranking candidate therapies, optimizing lead compounds, and understanding mechanism of action.

IC-50: The concentration of compound required to inhibit a specific biological or enzymatic activity by 50%, indicating potency and efficacy.

Ki: The equilibrium constant for inhibitor binding to its target, reflecting binding affinity and selectivity.

MIC: The minimum inhibitory concentration needed to prevent visible growth of microorganisms, used to assess antimicrobial or antiproliferative activity.

Recommended In Vitro Efficacy Tests

Abl Proto-Oncogene 1, Non-Receptor Tyrosine Kinase

Abl Proto-Oncogene 1, Non-Receptor Tyrosine Kinase (ABL1) is implicated in abnormal cell signaling in Systemic Mastocytosis, contributing to uncontrolled mast cell proliferation. Testing ABL1 activity is vital for identifying therapeutic targets and monitoring drug efficacy. Key methods include PCR, sequencing, and kinase assays. Main parameters assessed are ABL1 gene mutations, expression levels, and kinase activity, enabling precise characterization for drug development and patient stratification.

Atp Binding Cassette Subfamily B Member 1

Atp Binding Cassette Subfamily B Member 1 (ABCB1) mediates drug efflux, impacting therapeutic response in Systemic Mastocytosis. Testing ABCB1 activity is essential to evaluate drug resistance and optimize candidate selection. The Rhodamine 123 accumulation assay quantifies ABCB1 function by measuring intracellular dye retention. Minimum inhibitory concentration (MIC) is the key parameter assessed, guiding effective drug development and dosing strategies for Systemic Mastocytosis therapies.

Bruton Tyrosine Kinase

Bruton Tyrosine Kinase (BTK) is implicated in the abnormal mast cell signaling seen in Systemic Mastocytosis. BTK testing is crucial for drug development, enabling evaluation of inhibitor efficacy. Key assay methods include ATP, FRET, fluorescent, chemiluminescent, BRET, and HTRF assays, which accurately measure BTK activity. Main parameters assessed are IC-50 and Ki, providing quantitative insights into drug potency and binding affinity.

Fms Related Receptor Tyrosine Kinase 3

Fms Related Receptor Tyrosine Kinase 3 (FLT3) is implicated in the proliferation and survival of mast cells in Systemic Mastocytosis. FLT3 testing is crucial for identifying mutations and evaluating drug responses during drug development. Key methods include PCR-based mutation analysis, flow cytometry, and kinase activity assays. Main parameters assessed are FLT3 mutation status, expression levels, and downstream signaling activity, enabling targeted therapy optimization.

Kit Proto-Oncogene, Receptor Tyrosine Kinase

The Kit Proto-Oncogene, Receptor Tyrosine Kinase (KIT) is frequently mutated in systemic mastocytosis, driving abnormal mast cell proliferation. KIT testing is crucial for diagnosis, prognosis, and targeted drug development. Key methods include PCR, Sanger sequencing, and next-generation sequencing to detect KIT mutations (especially D816V). Main parameters assessed are mutation type, allele frequency, and mutation burden, guiding therapeutic strategies and clinical trial eligibility.

Lyn Proto-Oncogene, Src Family Tyrosine Kinase

The Lyn Proto-Oncogene, a Src Family Tyrosine Kinase, regulates mast cell activation and proliferation, contributing to Systemic Mastocytosis pathogenesis. Testing Lyn kinase activity or expression is vital for identifying drug targets and evaluating therapeutic efficacy. Key methods include Western blotting, kinase activity assays, and flow cytometry. Main parameters assessed are Lyn phosphorylation levels, total Lyn protein expression, and downstream signaling activation.

Mechanistic Target Of Rapamycin Kinase

Mechanistic Target Of Rapamycin (mTOR) kinase regulates mast cell proliferation and survival, contributing to Systemic Mastocytosis pathogenesis. mTOR kinase testing is crucial for evaluating drug efficacy targeting this pathway. Key methods include Western blot, ELISA, and kinase activity assays. Main parameters assessed are mTOR phosphorylation status, downstream signaling (e.g., p70S6K, 4EBP1), and cellular proliferation rates, enabling precise assessment of candidate therapeutics' impact on mTOR signaling.

Platelet Derived Growth Factor Receptor Alpha

Platelet Derived Growth Factor Receptor Alpha (PDGFRA) mutations can contribute to Systemic Mastocytosis pathogenesis and drug resistance. PDGFRA testing is vital for precise diagnosis, prognosis, and targeted therapy development. Key methods include PCR-based mutation analysis and next-generation sequencing. Main parameters assessed are mutation type, allele frequency, and expression levels, enabling informed drug candidate selection and patient stratification in clinical trials.