Cytoskeleton, Inc. has been a reliable source of compound screening services in the areas of pre-clinical drug development programs and early compound screening in primary HTS projects, as well as secondary screening and compound target validation. We have extensive experience in gene design and expression with an eye to produce highly purified biological active proteins. Our expertise in protein purification is the basis for the complimentary skill of assay design. We have produced many functional assays for kinesins (e.g. Eg5, CenPE, MKLP2), dynein (cytoplasmic), myosins (e.g. cardiac, smooth, skeletal and non-muscle isoforms), small G-proteins (e.g. Rho, Arf, Ral families), tubulins (e.g. tumor, plant and fungal origins) and actin binding proteins, 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. The hits from these assays have resulted in tubulin innibitors, microtubule inhibitors, kinesin inhibtiors, dynein inhibitors, myosin inhibitors, GEF inibitors and small G-protein inhibtiors. Other experience in antibody and ELISA technology 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, cardio-vascular research and neuroscience.
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Compound screening service with targets such as actins, tubulins and microtubules, molecular motors(kinesins and myosins), and small G-proteins like Ras p21, Arf1, Arf6, Cdc42, Rac1 and RhoA.
Protein purification with high biological activity and recombinant or native sources.
Enzyme assay design and development.
Antibody and ELISA development.
If you prefer to send an e-mail describing your requirements, please click here tservice@cytoskeleton.com or call USA-303-322-2254 x316.
Examples of compound screening services:
1. CenPE and Eg5 kinesin motor IC50 determinations using microtubule stimulated ATPase assays, with monthly deliveries of 10 to 30 compounds, results turnaround within one week.
2. Cancer tubulin (cell line and tumor derived) polymerization assays to determine IC50 of anti-tumor drugs, 4 to 22 compounds, results turnaround in with 3 weeks.
3. Tubulin/microtubule polymerization assays with single concentration screening, 10 to 160 compounds, turnaround within 2 weeks.
4. Calcium activated sarcomeric myosin S1 ATPase assays with tropomyosin/troponin coated F-actin (thin filaments) as substrate. Determine shift in pCa on addition of 24 compounds at two concentrations, turnaround within 4 weeks.
For more information click here.
Examples of protein purification are:
1. Cancer cell line tubulins are useful for developing drugs targeted toward cancer cells. We have isolated 1.0-5.0 mg of 90% pure tubulin from 50 g of cells.
2. Small G-protein Arf1 and Arf6 within 8 weeks to >85% purity, and suitable for 10,000 GEF assays (50 x 1mg).
We perform full quality control, which includes:
1. Full quantitation of yield, in mg per g tissue.
2. PAGE-SDS of purified proteins, stained with coomassie blue and scanned with a densitometer, to estimate purity.
3. Biological activity determination depending on the requirements of the customer.
For more information click here.
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.
If you prefer please send an e-mail describing your requirements to tservice@cytoskeleton.com
Available Screening modules
Screen name | Amount | Module Code | For more information click here |
Tubulin polymerization (col. 340nm) | 32 assays (1 IC50 + 1 Control IC50) | CDS001 | |
Tubulin polymerization (col. 340nm) | 32 assays (14 duplicates + 2 Controls + 2 blanks) | CDS002 | |
Tubulin polymerization (col. 340nm) | 96 assays (5 IC50 + 1 Control IC50) | CDS003 | |
Tubulin polymerization (col. 340nm) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS004 | |
Tubulin polymerization (col. 340nm) | 192 assays (10 IC50 + 2 Control IC50) | CDS005 | |
Tubulin polymerization (col. 340nm) | 192 assays (80 duplicates + 8 Controls + 8 blanks) | CDS006 | |
Tubulin polymerization (col. 340nm) | 384 assays (20 IC50 + 4 Control IC50) | CDS007 | |
Tubulin polymerization (col. 340nm) | 384 assays (160 duplicates + 16 Controls + 16 blanks) | CDS008 | |
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Tubulin polymerization (flr. 360nm/410nm) | 96 assays (5 IC50 + 1 Control IC50) | CDS009 | |
Tubulin polymerization (flr. 360nm/410nm) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS010 | |
Tubulin polymerization (flr. 360nm/410nm) | 192 assays (10 IC50 + 2 Control IC50 | CDS011 | |
Tubulin polymerization (flr. 360nm/410nm) | 192 assays (80 duplicates + 8 Controls + 8 blanks) | CDS012 | |
Tubulin polymerization (flr. 360nm/410nm) | 384 assays (20 IC50 + 4 Control IC50 | CDS013 | |
Tubulin polymerization (flr. 360nm/410nm) | 384 assays (160 duplicates + 16 Controls + 16 blanks) | CDS014 | |
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Tubulin polymerization (flr. 360nm/410nm) | 60 assays (5 IC50 + 1 Control IC50) | CDS015 | |
Tubulin polymerization (flr. 360nm/410nm) | 60 assays (28 duplicates + 4 Controls) | CDS016 | |
Tubulin polymerization (flr. 360nm/410nm) | 60 assays (5 IC50 + 1 Control IC50) | CDS017 | |
Tubulin polymerization (flr. 360nm/410nm) | 60 assays (28 duplicates + 4 Controls) | CDS018 | |
Tubulin polymerization (flr. 360nm/410nm) | 60 assays (5 IC50 + 1 Control IC50) | CDS019 | |
Tubulin polymerization (flr. 360nm/410nm) | 60 assays (28 duplicates + 4 Controls) | CDS020 | |
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Kinesin Motor assays (650nm) | 96-endpoint (5 IC50 + 1 Control IC50) | CDS050 | |
Kinesin Motor assays (650nm) | 96-endpoint (40 duplicates + 4 Controls + 4 blanks) | CDS051 | |
Kinesin Motor assays (360nm) | 96-kinetic (5 IC50 + 1 Control IC50) | CDS052 | |
Kinesin Motor assays (360nm) | 96-kinetic (40 duplicates + 4 Controls + 4 blanks) | CDS053 | |
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Myosin Motor assays (650nm) | 96-endpoint (5 IC50 + 1 Control IC50) | CDS054 | |
Myosin Motor assays (650nm) | 96-endpoint (40 duplicates + 4 Controls + 4 blanks) | CDS055 | |
Myosin Motor assays (360nm) | 96-kinetic (5 IC50 + 1 Control IC50) | CDS056 | |
Myosin Motor assays (360nm) | 96-kinetic (40 duplicates + 4 Controls + 4 blanks) | CDS057 | |
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| Sarcomere assay (360nm), contains myosin S1 fragment (motor domain) with tropomyosin / troponin(T/C/I) complex and actin filaments. | 96-endpoint (5 IC50 + 1 Control IC50) | CDS058 | |
Sarcomere assay (360nm), contains myosin S1 fragment (motor domain) with tropomyosin / troponin(T/C/I) complex and actin filaments. | 96-endpoint (40 duplicates + 4 Controls + 4 blanks) | CDS059 | |
| Sarcomere assay (360nm), contains myosin S1 fragment (motor domain) with tropomyosin / troponin(T/C/I) complex and actin filaments. | 96-kinetic (5 IC50 + 1 Control IC50) | CDS060 | |
| 96-kinetic (40 duplicates + 4 Controls + 4 blanks) | CDS061 | ||
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| FtsZ GTPase assay (650nm, endpoint) | 96-kinetic (5 IC50 + 1 Control IC50) | CDS250 | |
| FtsZ GTPase assay (650nm, endpoint) | 96-kinetic (40 duplicates + 4 Controls + 4 blanks) | CDS251 | |
| FtsZ GTPase assay (360nm, kinetic) | 96-kinetic (5 IC50 + 1 Control IC50) | CDS252 | |
| FtsZ GTPase assay (360nm, kinetic) | 96-kinetic (40 duplicates + 4 Controls + 4 blanks) | CDS253 | |
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Actin (skeletal muscle) polymerization | 96 assays (5 IC50s + 1 control) | CDS080 | |
Actin (skeletal muscle) polymerization | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS081 | |
Actin (non-muscle) polymerization | 96 assays (5 IC50s + 1 control) | CDS082 | |
Actin (non-muscle) polymerization | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS083 | |
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F-Actin Binding Spin-down Assays (skeletal muscle actin) | 24 assays (10 duplicates, four controls. | CDS090 | |
F-Actin Binding Spin-down Assays (non muscle actin) | 24 assays (10 duplicates, four controls. | CDS091 | |
GEF Assays | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS100 | |
GAP Assays | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS105 | |
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Tubulin ELISA (alpha) (total tubulin) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS200 | |
Tubulin ELISA (beta I, II, III, IV, V or VI) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS201 | |
Tubulin ELISA (acetylated alpha) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS202 | |
Tubulin ELISA (glutamylated) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS203 | |
Tubulin ELISA (glycinated) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS204 | |
Tubulin ELISA (tyrosinated) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS205 | |
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GLISA with extracts provided by Customer | 48 assays (20 duplicates + 4 Controls + 4 blanks) | CDS220 | |
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GLISA with extracts made at Cytoskeleton | 48 assays (20 duplicates + 4 Controls + 4 blanks) | CDS221 | |
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RhoA ELISA (total RhoA) | 96 assays (40 duplicates + 4 Controls + 4 blanks) | CDS222 |
Protein purification modules:
Type | Name | Example Protein | Amount | Mod. # | More information |
Recombinant protein | Small protein or protein domain (<30kDal) with gene provided by customer. | RhoA small G-protein | 10 x 100 µg | REC012 | |
Recombinant protein | Small protein or protein domain (<30kDal) with gene synthesis. | RhoA small G-protein | 10 x 100 µg | REC022 | |
Recombinant protein | Medium to large protein or protein domain (31-100 kDal) with gene synthesis. | CenPE kinesin | 10 x 100 µg | REC032 | |
Recombinant protein | Small protein or protein domain (<30kDal) with gene synthesis and expression in eukaryotic system. | RhoA small G-protein with lipid tail | 10 x 100 µg | REC041 | |
Recombinant protein | Medium to large protein or protein domain (31-100 kDal) with gene synthesis and expression in eukaryotic system. | Recombinant beta-Tubulin | 10 x 100 µg | REC042 | |
Recombinant protein | Medium to large protein or protein domain (31-100 kDal) with multiple gene synthesis and co-expression in eukaryotic system. (useful for protein complex production). | Recombinant alpha/beta- Tubulin heterodimer | 10 x 100 µg | REC043 | |
Native source protein | Cited protein purification | Arp2/3 complex (seven protein complex) | 10 x 100 µg | REC052 |