Protein Purification
Cytoskeleton, Inc. has been a reliable source of protein purification services in the areas of recombinant and native source proteins. We emphasize high purity and biological activity which enables the proteins to be used in many applications including diagnostic kits, pre-clinical drug development programs, early compound screening in primary HTS projects, and in ELISA formats. We also have extensive experience in gene design and expression with an eye to produce the correct orientation and solubility for particular applications. Many purifications use the unique skills and expertise in cytoskeletal and signal transductions proteins. Using this combined knowledge we have produced many functional assays for kinesins (e.g., Eg5, CenPE, MKLP2), myosins (e.g., cardiac, smooth, skeletal, and non-muscle isoforms, sarcomere assys), 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, profiling, 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.
A particular speciality of Cytoskeleton's design formats is the ability to rapidly create lyophilized formulations in appropriate daily use or weekly use amounts. This enables HTS use and reproducible daily performance. 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 protein biochemistry, cell biology, cancer biology, cardio-vascular research, and neuroscience.
For more details in protein purification services and modules click on the About tab above.
For rapidly available proteins i.e., less than two weeks delivery, see the table below.
If you have an immediate question please select from the buttons above or send an e-mail describing your requirements to tservice@cytoskeleton.com
Rapidly available proteins:
Name | Type and description | CS-item # | Stock status | |||
KIF22 Motor Domain | (5-378) His-Protein: wild-type (Human recombinant) | In stock | ||||
KIF18A Motor Domain | (1-374) His-Protein: wild-type (Human recombinant) | In stock | ||||
Fascin-1 Protein: wild-type | Human recombinant protein produced in a bacterial expression system. | In stock | ||||
Lysyl Oxidase | 32 KDal Form, Bovine Leg Tendon | In stock | ||||
SOS exchange domain (564-1049) protein | Human recombinant protein produced in a bacterial expression system. | In stock | ||||
RasGRF1 GEF Protein | Cdc25 Exchange Domain, aa1038-1270, MBP tag, human. | In stock | ||||
Tiam1 GEF Protein | DHPH Exchange Domain, aa1040-1406, MBP tag, human. | In stock | ||||
Vav1 GEF Protein | DHPHC1 Exchange Domain, Y174D mutant, aa168-522, 6xHis tag, human. | In stock | ||||
Vav2 GEF Protein | DH Exchange Domain, aa189-374, 6xHis tag, human | In stock | ||||
ARNO GEF domain wild-type protein | Human recombinant protein expressed in E. coli. | In stock | ||||
SOS2 exchange domain (563-1051) protein | Human recombinant protein produced in a bacterial expression system. |
| In stock | |||
RAPGEF5 Protein: Ras association and exchange domain (57– 580) | Wild-type, (Human recombinant, GST tagged). | CS-GE09
| In stock | |||
H-Ras p21 protein | Wild-type, 6xHis tag, human recombinant. | In stock | ||||
N-Ras p21 protein | Wild-type, 6xHis tag, human recombinant. | In stock, inquire for more information | ||||
R-Ras protein | Wild-type, 6xHis tag, human recombinant. | In stock, inquire for more information | ||||
K-Ras4B protein | Wild-type, 6xHis tag, human recombinant. | In stock | ||||
K-Ras4B G12V mutated protein | G12V mutant, 6xHis tag, human recombinant. | In stock | ||||
K-Ras4B G13D mutated protein | G13D mutant, 6xHis tag, human recombinant. | In stock | ||||
K-Ras4B G13S mutated protein | G13S mutant, 6xHis tag, human recombinant. | In stock, inquire for more information | ||||
K-Ras4B K128A mutated protein | K128A mutant, 6xHis tag, human recombinant. | In stock, inquire for more information | ||||
K-Ras4B Q61P mutated protein | Q61P mutant, 6xHis tag, human recombinant. | In stock, inquire for more information | ||||
K-Ras4B R135A mutated protein | R135A mutant, 6xHis tag, human recombinant. | |||||
K-Ras4B G12D+I36N mutated protein | G12D+I36N double mutant, 6xHis tag, human recombinant. | |||||
K-Ras4B G12D+D38A mutated protein | G12D+D38A double mutant, 6xHis tag, human recombinant. | |||||
K-Ras4B G12D mutated protein | G12D mutant, 6xHis tag, human recombinant. | Instock, inquire for more information | ||||
K-Ras4B G12C mutated protein | G12C mutant Human recombinant, 6xHis-tag. | CS-RS14
| Instock | |||
Arf1 protein | Wild-type, 6xHis tag, human recombinant. | CS-FR01 | In stock, inquire for more information | |||
Arf6 protein | Wild-type, 6xHis tag, human recombinant. | CS-FR06 | In stock, inquire for more information | |||
Purine Nucleoside Phosphorylase (PNP) | Recombinant protein 6xHis tagged. | In stock | ||||
KIF7 motor domain | Recombinant protein 6xHis tagged. | In stock | ||||
Dynein (cytoplasmic) | Native porcine brain source. | In stock | ||||
MKLP2 kinesin motor domain | Recombinant source, His tagged. | In stock | ||||
Heavy Meromyosin Protein (HMM Fragment) | Purified from Bovine heart tissue. | In stock | ||||
Myosin - cardiac S1 fragment | Native source - bovine heart. Suitable for soluble sarcomere assay. | In stock | ||||
Myosin - smooth muscle S1 fragment | Native source - chicken gizzard muscle. Suitable for soluble sarcomere assay. | In stock | ||||
Myosin - skeletal muscle S1 fragment | Native source - rabbit leg muscle. Suitable for soluble sarcomere assay. | In stock | ||||
| Tropomyosin / Tropomodulin complex | Native source - bovine heart. Suitable for soluble sarcomere assay. | CS-TT05 | In stock | |||
| Sarcomere, reconstituted | Cardiac sarcomere reconstituted in vitro, used for calcium activated myosin ATPase assays. | CS-TFC01 | In stock | |||
| Tubulin capture ELISA plate | ELISA plate coated with pan-tubulin sheep polyclonal antibody. | CS-ET01 | ||||
| Tubulin protein - sheep brain | Native source - sheep brain, > 99% pure. | CS-T234S | In stock, inquire for more information | |||
| Tubulin protein - Caki-1 human tumor | Native source - Human Caki-1 xenografts grown in mice. | CS-TM001 | In stock, inquire for more information | |||
| Tubulin beta-III isoform monoclonal antibody | Mouse mAb clone 17.C5. | CS-ABT03 | ||||
Pyrene Muscle Actin | Bovine cardiac muscle, > 99% pure. | CS-AP07 | In stock |
In this section we describe more about the process of producing a reliable source of protein, the deliverables and the timelines.
The process of producing your protein of interest:
Initial inquiry → Formulate scope and timeline → Generate quotation → Initiate project with a Purchase Order → Receive weekly updates → Receive product and datasheet.
The intial inquiry can be as simple as an e-mail to our Technical Service Department, or to fill out the Custom Service Department request form. We will get back to you with clarification questions and a proposal for scope (gene construct, purification tag, protein activity and purity), batch size, and timeline. If those are acceptable conditions then we will generate a formal quote which is useful for passing through your purchasing process and generating a Purchase Order (PO). Work will begin once the PO is received and we will give you a more detailed plan and keep you informed with weekly updates. At the end of the project proteins will be shipped in a desiccated chamber with a datasheet that indicates the activity and purity of the preparation.
Deliverables
Proteins are aimed to be delivered on time and within specifications. If there are unpredictable hold ups we will keep you informed and work hard to get the project back on track. Proteins are usually lyophilized for improved stability and this is included in the cost of the project. Products are usually packaged with two layers of desiccated protection to ensure stability, and a desiccating chamber is included for all custom orders. The datasheet contains details of the proteins activity and purity, usually we will characterize the protein with 2 or 3 enzyme parameters e.g., Vmax, Kcat or Kd, and these parameters are developed in agreement with your expectations.
Getting started
All we need from you is the protein's name, a quantity, and timeline, sent in an e-mail to tservice@cytoskeleton.com. We'll respond within 24 hours. We look forward to working with you to provide the required reagent in a suitable time frame.
Protein purification modules with deliverable and timelines:
Type | Name | Deliverables | Example Protein | Amount | Timeline (wk) | Mod. # |
Recombinant protein | Small protein or protein domain (< 30 kDal) with gene provided by customer. | Highly purified (> 90% pure), E. coli expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Datasheet and assay method. Activity is in line with published articles. | RhoA small G-protein | 10 x 100 µg | 4 | REC012 |
Recombinant protein | Small protein or protein domain (< 30 kDal) with gene synthesis. | Highly purified (> 90% pure), E. coli expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Expression plasmid containing gene. Datasheet and assay method. Activity is in line with published articles. | RhoA small G-protein | 10 x 100 µg | 8 | REC022 |
Recombinant protein | Medium to large protein or protein domain (31-100 kDal) with gene synthesis. | Highly purified (> 90% pure), E. coli expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Expression plasmid containing gene. Datasheet and assay method. Activity is in line with published articles. | CenPE kinesin | 10 x 100 µg | 10 | REC032 |
Recombinant protein | Small protein or protein domain (< 30 kDal) with gene synthesis and expression in eukaryotic system. | Highly purified (> 90% pure), CHO cell or Baculovirus/insect cell- expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Expression plasmid containing gene. Datasheet and assay method. Activity is in line with published articles. | RhoA small G-protein with lipid tail | 10 x 100 µg | 12 | REC041 |
Recombinant protein | Medium to large protein or protein domain (31-100 kDal) with gene synthesis and expression in eukaryotic system. | Highly purified (> 90% pure), CHO cell or Baculovirus/insect cell-expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Expression plasmid containing gene. Datasheet and assay method. Activity is in line with published articles. | Recombinant beta-Tubulin | 10 x 100 µg | 12 | 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). | Highly purified (> 90% pure), CHO cell or Baculovirus/insect cell-expressed, His-tagged active protein lyophilized in 10 x 100 µg aliquots (or more depending on yield). Expression plasmid containing gene. Datasheet and assay method. Activity is in line with published articles. | Recombinant alpha/beta- Tubulin heterodimer | 10 x 100 µg | 16 | REC043 |
Native source protein | Cited protein purification | Highly purified (> 80% pure) protein from tissue or eukaryotic cells, lyophilized in 10 x 100 µg aliquots (or more depending on yield). Datasheet and assay method. Activity is in line with published articles. | Arp2/3 complex (seven protein complex) | 10 x 100 µg | 4 to 20 | REC052 |
If you have any questions please send an e-mail describing your requirements to tservice@cytoskeleton.com.