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 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, 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 formts 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

SOS exchange domain (564-1049) protein

Human recombinant protein produced in a bacterial expression system

CS-GE02

In stock

RasGRF1 GEF Protein 

Cdc25 Exchange Domain, aa1038-1270, MBP tag, human.

CS-GE03

In stock

Tiam1 GEF Protein

DHPH Exchange Domain, aa1040-1406, MBP tag, human.

CS-GE04

In stock

Vav1 GEF Protein

DHPHC1 Exchange Domain, Y174D mutant, aa168-522, 6xHis tag, human.

CS-GE05

In stock

Vav2 GEF Protein

DH Exchange Domain, aa189-374, 6xHis tag, human.

CS-GE06

In stock

ARNO GEF domain wild type protein

Human recombinant protein expressed in E.coli

CS-GE07

In stock

SOS2 exchange domain (563-1051) protein

Human recombinant protein produced in a bacterial expression system

CS-GE08

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.

RS01

In stock

N-Ras p21 protein

Wild-type, 6xHis tag, human recombinant.

CS-RS02

Instock, inquire for more information

R-Ras protein

Wild-type, 6xHis tag, human recombinant.

CS-RS05

Instock, inquire for more information

K-Ras4B protein

Wild-type, 6xHis tag, human recombinant.

CS-RS03

In stock

K-Ras4B G12V mutated protein

G12V mutant, 6xHis tag, human recombinant.

CS-RS04

In stock

K-Ras4B G13D mutated protein

G13D mutant, 6xHis tag, human recombinant.

CS-RS06

In stock

K-Ras4B G13S mutated protein

G13S mutant, 6xHis tag, human recombinant.

CS-RS07

Instock, inquire for more information

K-Ras4B K128A mutated protein

K128A mutant, 6xHis tag, human recombinant.

CS-RS08

Instock, inquire for more information

K-Ras4B Q61P mutated protein

Q61P mutant, 6xHis tag, human recombinant.

CS-RS09

Instock, inquire for more information

K-Ras4B R135A mutated protein

R135A mutant, 6xHis tag, human recombinant.

CS-RS10

Inquire

K-Ras4B G12D+I36N mutated protein

G12D+I36N double mutant, 6xHis tag, human recombinant.

CS-RS11

Inquire

K-Ras4B G12D+D38A mutated protein

G12D+D38A double mutant, 6xHis tag, human recombinant.

CS-RS12

Inquire

K-Ras4B G12D mutated protein

G12D mutant, 6xHis tag, human recombinant.

CS-RS13

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

Instock, inquire for more information

Arf6 protein

wild-type, 6xHis tag, human recombinant.

CS-FR06

Instock, inquire for more information

Purine Nucleoside Phosphorylase (PNP)

 

Recombinant protein 6xHis tagged

 

ELP03

 

 In stock

KIF7 motor domain

 

Recombinant protein 6xHis tagged

 

CS-KF51

 

 In stock

Dynein (cytoplasmic)

 

Native porcine brain source.

 

CS-DN01

 

 In stock

MKLP2 kinesin motor domain

 

Recombinant source His tagged.

 

CS-MP05

 

 In stock

Heavy Meromyosin Protein (HMM Fragment)

 

Purified from Bovine heart tissue

 

CS-MH03

 

 In stock

Myosin - cardiac S1 fragment

 

Native source - bovine heart. Suitable for soluble sarcomere assay. 

 

CS-MYS03

 

 In stock

Myosin - smooth muscle S1 fragment

 

Native source - chicken gizzard muscle. Suitable for soluble sarcomere assay. 

 

CS-MYS05

 

 In stock

Myosin - skeletal muscle S1 fragment

 

Native source - rabbit leg muscle. Suitable for soluble sarcomere assay. 

 

CS-MYS04

 

 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  

 Inquire

Tubulin protein - sheep brain Native source - sheep brain, >99% pure. CS-T234S 

 Instock, inquire for more information

Tubulin protein - Caki-1 human tumor Native source - Human Caki-1 xenografts grown in mice.CS-TM001 

 Instock, inquire for more information

Tubulin beta III isoform monoclonal antibody  Mouse MAb clone 17.C5  CS-ABT03 

 Inquire

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 attached form (click for Form). We will get back to you with some 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). When we receive the PO the work can start, we will give you a more detailed plan and keep you informed with weekly updates. At the end of the project we will ship the proteins in a desiccated chamber and provide a datasheet which 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, send this information in an e-mail to tservice@cytoskeleton.com and 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 (<30kDal) 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 (<30kDal) 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 contianing 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 (<30kDal) 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.