Kits and Assays FAQs
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 vivo data 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.