Assay Development

Cytoskeleton, Inc. has been a reliable source of assay design and development services for the past 20 years. We emphasize highly accurate and functional enzyme formats using highly purified and biologically activity proteins which enables the assays to reproduce the enzymatic parameters in the published literature, i.e., similar Kd, Kcat, Vmax, pKa or IC50 values, and dose-responses.

A particular speciality of Cytoskeleton's design formats is the ability to rapidly formulate lyosphilized formulations in appropriate daily use or weekly use amounts. This enables HTS use and reproducible daily performance.

We also have extensive experience in gene design and expression with an eye to produce the correct orientation and solubility for particular applications.

Many assays use the unique skills and expertise in cytoskeletal and signal transductions proteins that no other company can match. Using this combined knowledge we have produced many functional assays for GTP exchange factors (GEFs) (e.g., Dbs, LARG, SOS1, Tiam1, Vav1, and Vav2), kinesins (e.g., Eg5, CenPE, MKLP2), myosins (e.g., cardiac, smooth, skeletal and non-muscle isoforms, sarcomere assays), small G-proteins (e.g., Rho, Arf, Ral families), tubulins (e.g., tumor, plant, and fungal origins), tubulin binding proteins (e.g., tau and MAPs) and actin binding proteins (e.g., cofilin, profilin, gelsolin), many of them are multi-protein assays that might have protein complexes of 3 or more subunits e.g., a soluble sarcomere format and the Arp2/3 complex based assay.

There are a set of defined modules that allow one to chose the desired package for your application, see below for module details. Other experience in antibody and ELISA technology and protein synthesis complements the cytoskeletal and signal transduction focus. We support all of our services with a dedicated technical services department and years of laboratory experience in the fields of cell biology, cancer biology, and neuroscience.

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Examples of assay development are:

1. Calcium activated sarcomeric myosin S1 ATPase assays with tropomyosin/troponin coated F-actin (thin filaments) as substrate, assay is able to determine shift in pCa on addition compound with a CV = 5.6%.

2. Plant cell tubulin polymerization assay within 8 weeks. Microliter scale (10 µl) and with a CV = 14%.

3. Small G-protein Arf1 and Arf6 GEF assays within 4 weeks with CV = 7.8%.

4. Microtubule stablization development for kinesin motor drug screen, resulting in one week room temperature stable preparation of microtubules, with intra assay CV = 7.5% and day-to-day variation CV = 8%. For more information click here.