Kits & Assays

Cytoskeleton's Biochem Kits™ are comprehensive kits for assaying different aspects of signal transduction processes and cytoskeletal biochemistry.  These kits help scientists produce publication quality data in a short period of time (Click citation tab above for examples).  These kits come with all the reagents needed for the assay as well as detailed instruction on how to use them, so you will be ready to perform experiments as soon as the kits arrives!

 

For more information about our Biochem Kits please click the About tab above.

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About Cytoskeleton's Biochem™ kits

Biochem™ kits are an extremely quick way to become familiar with some of the most useful in vitro and in vivo assays in signal transduction and cytoskeletal research. Each kit is carefully quality controlled and very user friendly and will therefore save time and money even for experienced cytoskeletal researchers. Cytoskeleton provides Biochem kits for actin, tubulin, intermediate filament, small G-protein, signal transduction and molecular motor research.

If you are new to the field of cytoskeletal research, the Biochem™ kits will prove to be an invaluable introductory aid, providing many useful technical tips. For example, if you are new to the field of microtubule research, you may not know that tubulin requires GTP, Mg2+ and >50 mM PIPES for stability and that taxol (a microtubule stabilizing drug) added too quickly will cause aberrant tubulin arrangements to form. If either of these points are overlooked, the results could lead to inappropriate experimental interpretation.Biochem™ kits are designed to overcome this bottleneck in research.

As an example: If you wanted to know whether a specific protein or compound interacts with actin filaments and/or monomer, you could order one of several Biochem kits specializing in actin research:

1) Actin binding protein assay Biochem™ kit (Cat. # BK001 or BK013): This kit provides clear instructions and all the reagents and controls necessary to determine the ability of a given protein to bind to actin filaments. It should be noted that in some cases actin binding proteins have a low affinity for actin filaments. Therefore, it is wise to couple this assay with the Actin polymerization assay Biochem™ kit in order to obtain a more complete profile of your potential actin binding protein.

2) Actin polymerization assay Biochem™ kit (Cat. # BK003): This kit uses modified actin to follow polymerization kinetics. There are many types of actin binding proteins, including nucleating, capping, severing, side-binding and monomer sequestering actin binding proteins. All of these can be characterized by their effect on actin polymerization. Cytoskeleton has adapted this powerful assay into a user friendly Biochem™ kit format.

Biochem™ kits are also a great way to optimize your research time and money. When you buy a Biochem™ kit, you obtain all the necessary proteins, buffers, controls and instructions that you require to perform your research with the highest degree of confidence. Furthermore, Cytoskeleton Inc guarantees that the cost of each kit is substantially less than the sum of the component parts purchased separately.

Many publications cite the use of Cytoskeleton's kits in the Materials and Methods section of papers. Usually the citation is associated with a particular result in the form of a graph or image that helps the authors present their findings. This indicates the utility of the Kits to produce publication quality data in a short timeframe thus helping improve the productivity of your efforts. Example citations for different classes of kit, more citations are available on individual product pages.

Actin Kits, e.g. Actin Polymerization Assay Kit (Cat. # BK003)

Takamiya, R., Takahashi, M., Park, Y. S., Tawara, Y., Fujiwara, N., Miyamoto, Y., Gu, J., Suzuki, K. and Taniguchi, N. (2005). Overexpression of mutated Cu,Zn-SOD in neuroblastoma cells results in cytoskeletal change. Am. J. Physiol. 288, C253-259.

Kumar, N., Tomar, A., Parrill, A. L. and Khurana, S. (2004). Functional dissection and molecular characterization of calcium-sensitive actin-capping and actin-depolymerizing sites in villin. J. Biol. Chem. 279, 45036-45046.

Small G-protein kits, e.g. GLISA Activation Assays

Alvarez et al. (2010). Failure of Bay K 8644 to induce RhoA kinase-dependent calcium sensitization in rabbit blood vessels. British J of Pharmacology 160 ,1326-37.

Heckman-Stoddard et al. (2009). Haploinsufficiency for p190B RhoGAP inhibits MMTV-Neu tumor progression. Breast Cancer Research 11 ,http://breast-cancer-research.com/content/11/4/R61.

Tubulin kits, e.g. Tubulin polymerization assay (Cat. # BK006P)

O'Boyle NM, Carr M, Greene LM, Bergin O, Nathwani SM, McCabe T, Lloyd DG, Zisterer DM, Meegan MJ. (2010). Synthesis and Evaluation of Azetidinone Analogues of Combretastatin A-4 as Tubulin Targeting Agents.. J Med Chem.

Jacob Kushkuley, Walter K. H. Chan, Sangmook Lee, Joel Eyer, Jean-Francois Leterrier, Franck Letournel and Thomas B. Shea (2009). Neurofilament cross-bridging competes with kinesin-dependent association of neurofilaments with microtubules. J Cell Science 122 ,3579-86.

Protein Assays, e.g. Advanced Protein Assay (Cat. # ADV01)

Bachran C, Schneider S, Riese SB, Bachran D, Urban R, Schellmann N, Zahn C, Sutherland M, Fuchs H. (2010). A lysine-free mutant of epidermal growth factor as targeting moiety of a targeted toxin. Life Sci.

 

Honnappa, S., Cutting, B., Jahnke, W., Seelig, J. and Steinmetz, M. O. (2003). Thermodynamics of the Op18/stathmin-tubulin interaction. J. Biol. Chem. 278, 38926-38934.

Question 1:  How can Cytoskeleton’s Biochem Kits help me in my research?

Question 2:  How can I use the Biochem Kits to investigate whether my protein interacts with the cytoskeleton or its regulatory components?

 


Question 1: How can Cytoskeleton’s Biochem Kits help me in my research?

Answer 1:Cytoskeleton's Biochem Kits are an extremely quick way to become familiar with some of the most useful in vitro and in vivo assays for assaying different aspects of cytoskeletal biochemistry and signal transduction. These kits come with all the reagents needed for your assay as well as detailed instruction on how to use them, so you will be ready to do your assays as soon as you have the kits!  Each kit is carefully quality controlled and very user friendly and will therefore save time and money even for experienced cytoskeletal researchers.  Furthermore, Cytoskeleton, Inc. guarantees that the cost of each kit is substantially less than the sum of the component parts purchased separately.  Cytoskeleton, Inc. provides Biochem Kits for actin, tubulin, G-protein activation, G-protein signal transduction and molecular motor research.

If you are new to the field of cytoskeletal research, the Biochem Kits will prove to be an invaluable introductory aid, providing many useful technical tips. For example, if you are new to the field of microtubule research, you may not know that tubulin requires GTP, Mg2+ and >50 mM PIPES for stability and that taxol (a microtubule stabilizing drug) added too quickly will cause aberrant tubulin arrangements to form. If any of these points are overlooked, the results could lead to inappropriate experimental interpretation.Biochem kits are designed to overcome this bottleneck in research.

Question 2: How can I use the Biochem Kits to investigate whether my protein interacts with the cytoskeleton or its regulatory components?

Answer 2:Cytoskeleton, Inc. has a variety of kits that can help the researcher examine how recombinant proteins, drugs or even cell lysates either interact and/or affect the function of cytoskeletal proteins including monomers and polymers of actin and tubulin. 

Some examples of kits specifically designed for this purpose include the actin binding protein assay Biochem Kit (Cat. # BK001 or BK013) and the tubulin binding protein assay Biochem Kit (Cat.  # BK029). 

Actin binding protein assay Biochem kit (Cat.  # BK001 or BK013): This kit provides clear instructions and all the reagents and controls necessary to determine the ability of a given protein to bind to actin monomers versus actin filaments.  F-actin binding can be measured by using a spin down assay where centrifugation is used to separate F-actin from G-actin by differential sedimentation.  This kit can be used to determine (i) whether a test protein binds F-actin or affects the equilibrium between G-actin and F-actin, (ii) whether a test protein has G-actin sequestering or F-actin polymerization enhancing activity or (iii) whether a test protein of interest can bundle F-actin.  We suggest coupling this assay with the Actin polymerization assay Biochem kit in order to obtain a more complete profile of your potential actin binding protein.

Tubulin binding protein assay Biochem Kit(Cat. # BK029):This kit provides clear instructions and all the reagents and controls necessary to determine the ability of a given protein to bind to tubulin monomers versus tubulin polymers (microtubules).  Tubulin binding can be measured by using a spin down assay where centrifugation is used to separate microtubules from tubulin monomers by differential sedimentation.  This kit can be used to: (i) identify novel microtubule associated proteins (MAPs), (ii) confirm in vivodata suggesting a given protein is a MAP, (iii) characterize MAPs, (iv) identify/characterize MAP regulating proteins or (v) identify/characterize compounds that inhibit MAP binding to microtubules.  We suggest coupling this assay with the tubulin polymerization assay Biochem Kit (Cat. # BK006P or BK011P) in order to obtain a more complete profile of your potential tubulin binding protein. 

 

Actin polymerization assay Biochem kit (Cat. # BK003): This kit uses modified actin to follow polymerization kinetics. There are many types of actin binding proteins, including nucleating, capping, severing, side-binding and monomer sequestering actin binding proteins. All of these can be characterized by their effect on one or more of the three stages of actin polymerization: nucleation, growth and steady-state equilibrium.  A compound’s effects on depolymerization can also be evaluated. Cytoskeleton has adapted this powerful assay into a user friendly Biochem kit format.

Tubulin polymerization assay Biochem kit (Cat. # BK006P or BK001P): We offer both absorbance-based and fluorescence-based tubulin polymerization kits to follow polymerization kinetics.  The absorbance-based kit (Cat. # BK006P) takes advantage of the fact that light is scattered by microtubules to an extent that is proportional to the concentration of microtubule polymer.  The fluorescence-based kit (Cat. # BK011P) measures polymerization as a function of fluorescence enhancement following the incorporation of a fluorescent reporter into microtubules as polymerization occurs.  There are many proteins and drugs that either enhance or inhibit polymerization or depolymerization.  These compounds can be characterized by their effect on one or more of the three stages of tubulin polymerization: nucleation, growth, and steady-state equilibrium.  A compound’s effects on depolymerization can also be evaluated. Cytoskeleton has adapted these powerful assays into user friendly Biochem Kit formats.

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