Gliding Assay to Analyze Microtubule-based Motor Protein Dynamics.

Gliding Assay to Analyze Microtubule-based Motor Protein Dynamics.

Bio-protocol (2017-04-05)

Albert Shim, Gin Tezuka, Luke Kupcha, Li Tao

ABSTRACT

The purpose of this protocol is to provide an updated method of performing microtubule gliding assays and visualizing it using fluorescence microscopy.

MATERIALS

Product Number

Brand

Product Description

L2884

Sigma-Aldrich

Leupeptin, microbial, ≥90% (HPLC)

A3428

Sigma-Aldrich

Aprotinin from bovine lung, lyophilized powder, 3-8 TIU/mg solid, BioReagent, suitable for cell culture

E3889

Sigma-Aldrich

Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, Molecular Biology, ≥97.0%

G7141

Sigma-Aldrich

Glucose Oxidase from Aspergillus niger, Type X-S, lyophilized powder, 100,000-250,000 units/g solid (without added oxygen)

D9779

Sigma-Aldrich

DL-Dithiothreitol, Molecular Biology, ≥98% (HPLC), ≥99% (titration)

T6522

Sigma-Aldrich

Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture

G8877

Sigma-Aldrich

Guanosine 5′-triphosphate sodium salt hydrate, ≥95% (HPLC), powder

P6757

Sigma-Aldrich

PIPES, ≥99% (titration)

P5318

Sigma-Aldrich

Pepstatin A, microbial, ≥90% (HPLC)

P7626

Sigma-Aldrich

Phenylmethanesulfonyl fluoride, ≥98.5% (GC)

P5405

Sigma-Aldrich

Potassium chloride, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%

T1503

Sigma-Aldrich

Trizma^® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline

C1345

Sigma-Aldrich

Catalase from bovine liver, powder, suitable for cell culture, 2,000-5,000 units/mg protein

C7078

Sigma-Aldrich

Casein from bovine milk, technical grade

M4880

Sigma-Aldrich

Magnesium chloride, suitable for insect cell culture, BioReagent, ≥97.0%

E3876

Sigma-Aldrich

N-Ethylmaleimide, crystalline, ≥98% (HPLC)

A6419

Sigma-Aldrich

Adenosine 5′-triphosphate disodium salt hydrate, microbial, BioReagent, suitable for cell culture, ≥99% (HPLC)

P5368

Sigma-Aldrich

Phosphate buffered saline, BioPerformance Certified, pH 7.4

D9434

Sigma-Aldrich

Dextrose, 97.5-102.0% anhydrous basis, meets EP, BP, JP, USP testing specifications