Tubulin polymerization assay using >99% pure tubulin, fluorescence based (BK011P)

Tubulin polymerization HTS assay using >99% pure tubulin, fluorescence based (BK011P)

$0.00

SKU

BK011

Datasheet

MSDS

Amount

96 assays (Cat. # BK011P) + $968.00

Qty:

Details

Product Uses Include

Cost effective high throughput screen for anti-cancer compounds.
Basic research to measure compound IC50s and specificity for tubulin.
Screening proteins for effects on tubulin polymerization activity.
Teaching aid for undergraduate/graduate class in pharmacology.

Introduction
This assay is an economical one step procedure for determining the effects of drugs or proteins on tubulin polymerization. It is an adaptation of an assay originally described by Bonne, D. et al. (1). Polymerization is followed by fluorescence enhancement due to the incorporation of a fluorescent reporter into microtubules as polymerization occurs. The standard assay uses neuronal tubulin (Cat. # T240), which generates a polymerization curve representing the three phases of microtubule formation; namely nucleation, growth and steady state equilibrium. Other tubulins, such as HeLa cell tubulin (Cat. # H001) can also be used in this assay. The low volume per assay of 50 µl and the low tubulin concentrations, 2 mg/ml final concentration (100 µg per assay), makes this an ideal choice for studying the more expensive cancer cell tubulin reagents and for high throughput applications.

The classic tubulin polymerization assay uses absorbance readings at 340 nm to follow microtubule formation. It is based upon the fact that light is scattered by microtubules to an extent that is proportional to the concentration of microtubule polymer. This assay is offered by Cytoskeleton, Inc. (Cat. # BK006P). The fluorescence based assay has been compared to the absorbance based format and the comparisons are given in Table 1 below. For help in selecting the best assay format for your needs, contact tservice@cytoskeleton.com.

Click Here for Detailed Methods

Table 1. Comparison of Fluorescence versus Absorbance Based Polymerization Assays

Assay Characteristics	Absorbance Assay	Fluorescence Assay
Tubulin used per assay	300 µg	100 µg
Volume of reaction	100 µl	50 µl
Signal to noise ratio (S/N)	2	4
Coefficient of variation (cv)*	13%	11%
Paclitaxel EC50**	1 µM	1 µM
Vinblastine IC50 **	0.6 µM	0.6 µM
Possible problems	Glycerol in standard assay format may interfere with drug or protein binding. Assay conditions can easily be altered to test this.	Fluorescent reporter may interfere with drug or protein binding.
: Duplicate samples *: Under standard assay conditions. Conditions can be optimized for polymerization enhancers or inhibitors.

Kit contents
The kit contains sufficient material for 96 assays in 50 µl format. The following components are included:

>99% pure tubulin (Cat. # T240)
General tubulin buffer with fluorescent reporter
Microtubule glycerol buffer (Cat. # BST05)
GTP solution (Cat. # BST06)
Paclitaxel positive control (Cat. # TXD01)
Half area 96-well plate. Black, flat bottom
Manual with detailed protocols and extensive troubleshooting guide

Equipment needed

Tempererature controlled 96-well plate fluorimeter equipped with filters for exitation at 340-360 nm and emission at 420-450 nm

Example results
Compounds or proteins that interact with tubulin will often alter one or more of the characteristic phases of polymerization. For example, Figure 1 shows the effect of adding the anti-mitotic drug paclitaxel to the standard polymerization reaction. A 3 µM concentration of paclitaxel eliminates the nucleation phase and enhances the Vmax of the growth phase. Thus, one application of this assay is the identification of novel anti-mitotics. Figure 1 also shows the effect of adding the microtubule destabilizing drug, vinblastine. At 3 µM final concentration, vinblastine causes a drastic decrease in Vmax and reduction in final polymer mass.

Figure 1. Tubulin polymerization using the fluorescence based tubulin polymerization assay (BK011P). Tubulin was incubated alone (Control), with Paclitaxel or Vinblastine. Each condition was tested in duplicate. Polymerization was measured by excitation at 360 nm and emission at 420 nm. The three Phases of tubulin polymerization are marked for the control polymerization curve; I: nucleation, II: growth, III: steady state equillibrium.

References

Bonne, D., Heusele, C., Simon, C., and Pantaloni, D. (1985). 4’, 6-Diamidino-2-phenylindole, a fluorescent probe for tubulin and mictrotubules. J. Biol. Chem. 260, 2819-2825.

About

For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com

For our Tubulin Polymerization (Fluorescence) Excel Template please download here.
For our IC50 from Vmax Polymerization Excel Template please download here.

Citations

Author	Title	Journal	Year	Article Link
Kumar N, Manoj et al.	Benzothiazole-[1,2,3]triazolo[5,1-a]isoindoles: Synthesis, anticancer activity, bioavailability and in silico studies against Gama-Tubulin protein	Journal of Molecular Structure	2022	ISSN 0022-2860
Yao, Yongfang et al.	Angiogenesis and anti-leukaemia activity of novel indole derivatives as potent colchicine binding site inhibitors	Journal of Enzyme Inhibition and Medicinal Chemistry	2022	ISSN 1475-6374
Wang, Zhan et al.	Design, synthesis and biological evaluation of colchicine glycoconjugates as tubulin polymerization inhibitors	Bioorganic and Medicinal Chemistry	2022	ISSN 1464-3391
Ma, Juan et al.	Discovery of Novel 3,4-Dihydro-2(1H)-Quinolinone Sulfonamide Derivatives as New Tubulin Polymerization Inhibitors with Anti-Cancer Activity	Molecules	2022	ISSN 1420-3049
Würtz, Martin et al.	Modular assembly of the principal microtubule nucleator γ-TuRC	Nature Communications	2022	ISSN 2041-1723
Liu, Chao et al.	Synthesis and biological evaluation of BU-4664L derivatives as potential anticancer agents	Bioorganic and Medicinal Chemistry Letters	2022	ISSN 1464-3405
Wang, Guangcheng et al.	Design, synthesis and biological evaluation of novel 2-phenyl-4,5,6,7-tetrahydro-1H-indole derivatives as potential anticancer agents and tubulin polymerization inhibitors	Arabian Journal of Chemistry	2022	ISSN 1878-5352
Patel, Onisha et al.	Structural basis for small molecule targeting of Doublecortin Like Kinase 1 with DCLK1-IN-1	Communications Biology	2021	ISSN 2399-3642
Fareed, Momen R. et al.	New multi-targeted antiproliferative agents: Design and synthesis of ic261-based oxindoles as potential tubulin, ck1 and egfr inhibitors	Pharmaceuticals	2021	ISSN 1424-8247
Rahimzadeh Oskuei, Sara et al.	Design, synthesis and biological evaluation of novel imidazole-chalcone derivatives as potential anticancer agents and tubulin polymerization inhibitors	Bioorganic Chemistry	2021	ISSN 1090-2120
Liang, Dong et al.	Identification of anthelmintic parbendazole as a therapeutic molecule for HNSCC through connectivity map-based drug repositioning	Acta Pharmaceutica Sinica B	2021	ISSN 2211-3843
Sun, Chiao Yin et al.	LMBD1 protein participates in cell mitosis by regulating microtubule assembly	Biochemical Journal	2021	ISSN 1470-8728
Wang, Chao et al.	Design, synthesis and biological evaluation of 1-Aryl-5-(4-arylpiperazine-1-carbonyl)-1H-tetrazols as novel microtubule destabilizers	Journal of Enzyme Inhibition and Medicinal Chemistry	2021	ISSN 1475-6374
Würtz, Martin et al.	Reconstitution of the recombinant human γ-tubulin ring complex	Open Biology	2021	ISSN 2046-2441
Grohmann, Christoph et al.	Preclinical small molecule WEHI-7326 overcomes drug resistance and elicits response in patient-derived xenograft models of human treatment-refractory tumors	Cell Death and Disease	2021	ISSN 2041-4889
Yang, Mei et al.	C118P, a novel microtubule inhibitor with anti-angiogenic and vascular disrupting activities, exerts anti-tumor effects against hepatocellular carcinoma	Biochemical Pharmacology	2021	ISSN 0006--2952
	Lead optimization of novel quinolone chalcone compounds by a structure-activity relationship (SAR) study to increase efficacy and metabolic stability		2021	Article Link
Shawky, Ahmed M. et al.	Novel pyrrolizines bearing 3,4,5-trimethoxyphenyl moiety: design, synthesis, molecular docking, and biological evaluation as potential multi-target cytotoxic agents	Journal of Enzyme Inhibition and Medicinal Chemistry	2021	ISSN 1475-6374
Woo, Jung AA et al.	β-arrestin1 promotes tauopathy by transducing GPCR signaling, disrupting microtubules and autophagy	Life science alliance	2021	ISSN 2575--1077
Liu, Qian et al.	Identification of a lathyrane-type diterpenoid EM-E-11-4 as a novel paclitaxel resistance reversing agent with multiple mechanisms of action	Aging	2020	ISSN 1945-4589
Abdel-Rahman, Somaya A. et al.	Cyclohepta[ b]thiophenes as Potential Antiproliferative Agents: Design, Synthesis, in Vitro, and in Vivo Anticancer Evaluation	ACS Pharmacology and Translational Science	2020	ISSN 2575-9108
Barnes, Natalie G. et al.	A 2-step synthesis of Combretastatin A-4 and derivatives as potent tubulin assembly inhibitors	Bioorganic and Medicinal Chemistry	2020	ISSN 1464-3391
Palumbo, Valeria et al.	Drosophila Morgana is an Hsp90-interacting protein with a direct role in microtubule polymerization	Journal of Cell Science	2020	ISSN 1477-9137
Mirzaei, Salimeh et al.	Synthesis, structure-activity relationship and molecular docking studies of novel quinoline-chalcone hybrids as potential anticancer agents and tubulin inhibitors	Journal of Molecular Structure	2020	ISSN 0022-2860
Chen, Shih Yin et al.	Exosomal 2′,3′-CNP from mesenchymal stem cells promotes hippocampus CA1 neurogenesis/neuritogenesis and contributes to rescue of cognition/learning deficiencies of damaged brain	Stem Cells Translational Medicine	2020	ISSN 2157-6580
Song, Ming Yu et al.	Exploring diverse-ring analogues on combretastatin a4 (Ca-4) olefin as microtubule-targeting agents	International Journal of Molecular Sciences	2020	ISSN 1422-0067
Ullah, Imran et al.	An Antiparasitic Compound from the Medicines for Malaria Venture Pathogen Box Promotes Leishmania Tubulin Polymerization	ACS Infectious Diseases	2020	ISSN 2373-8227
Tariq, Ammarah et al.	In vitro reconstitution of branching microtubule nucleation	eLife	2020	ISSN 2050-084X
Zhernov, Ilia et al.	Intrinsically Disordered Domain of Kinesin-3 Kif14 Enables Unique Functional Diversity	Current Biology	2020	ISSN 1879-0445
Zdioruk, Mykola et al.	A new inhibitor of tubulin polymerization kills multiple cancer cell types and reveals p21-mediated mechanism determining cell death after mitotic catastrophe	Cancers	2020	ISSN 2072-6694
Skidmore, Lillian et al.	ARX788, a site-specific anti-HER2 antibody–drug conjugate, demonstrates potent and selective activity in HER2-low and T-DM1–resistant breast and gastric cancers	Molecular Cancer Therapeutics	2020	ISSN 1538-8514
Wang, Yanming et al.	Antibody-drug conjugate using ionized CYS-linker-mmae as the potent payload shows optimal therapeutic safety	Cancers	2020	ISSN 2072-6694
Baker, Stacey J. et al.	A Contaminant Impurity, Not Rigosertib, Is a Tubulin Binding Agent	Molecular Cell	2020	ISSN 1097-4164
Morita, Ken et al.	Allosteric Activators of Protein Phosphatase 2A Display Broad Antitumor Activity Mediated by Dephosphorylation of MYBL2	Cell	2020	ISSN 1097-4172
Dillon, Gregory M. et al.	Acute inhibition of the CNS-specific kinase TTBK1 significantly lowers tau phosphorylation at several disease relevant sites	PLoS ONE	2020	ISSN 1932-6203
Zhai, Min'an et al.	3,5-Diaryl-1H-pyrazolo[3,4-b]pyridines as potent tubulin polymerization inhibitors: Rational design, synthesis and biological evaluation	European Journal of Medicinal Chemistry	2019	ISSN 1768-3254
Su, Wenhui et al.	Cdc42 is involved in NC1 peptide-regulated BTB dynamics through actin and microtubule cytoskeletal reorganization	FASEB journal : official publication of the Federation of American Societies for Experimental Biology	2019	ISSN 1530-6860
Wang, Yanming et al.	Novel silyl ether-based acid-cleavable antibody-MMAE conjugates with appropriate stability and efficacy	Cancers	2019	ISSN 2072-6694
Cui, Ying Jie et al.	Synthesis of novel pyrazole derivatives and their tumor cell growth inhibitory activity	Molecules	2019	ISSN 1420-3049
Shaik, Thoukhir B. et al.	Evaluation of Anticancer and Anti-Mitotic Properties of Quinazoline and Quinazolino-Benzothiadiazine Derivatives	Anti-Cancer Agents in Medicinal Chemistry	2019	ISSN 1871-5206
Sonawane, Vinay et al.	Cink4T, a quinazolinone-based dual inhibitor of Cdk4 and tubulin polymerization, identified via ligand-based virtual screening, for efficient anticancer therapy	European Journal of Medicinal Chemistry	2019	ISSN 1768-3254
Xia, Xiaoyu et al.	Leukemia Cell Cycle Chemical Profiling Identifies the G2-Phase Leukemia Specific Inhibitor Leusin-1	ACS Chemical Biology	2019	ISSN 1554-8937
Behbahani, Fatemeh Shaebani et al.	Synthesis and biological evaluation of novel benzo[c]acridine-diones as potential anticancer agents and tubulin polymerization inhibitors	Archiv der Pharmazie	2019	ISSN 1521-4184
Karimikia, Ehsan et al.	Colchicine-like β-acetamidoketones as inhibitors of microtubule polymerization: Design, synthesis and biological evaluation of in vitro anticancer activity	Iranian Journal of Basic Medical Sciences	2019	ISSN 2008-3874
Patterson, Jesse C. et al.	VISAGE Reveals a Targetable Mitotic Spindle Vulnerability in Cancer Cells	Cell Systems	2019	ISSN 2405-4720
Mao, Bai Ping et al.	CaMSAP2 is a microtubule minus-end targeting protein that regulates BTB dynamics through cytoskeletal organization	Endocrinology	2019	ISSN 1945-7170
Eberle-Singh, Jaime A. et al.	Effective delivery of a microtubule polymerization inhibitor synergizes with standard regimens in models of pancreatic ductal adenocarcinoma	Clinical Cancer Research	2019	ISSN 1557-3265
Lukinavičius, Gražvydas et al.	Fluorescent dyes and probes for super-resolution microscopy of microtubules and tracheoles in living cells and tissues	Chemical Science	2018	ISSN 2041-6539
Tian, Zhenhua et al.	Biological activity and interaction mechanism of the diketopiperazine derivatives as tubulin polymerization inhibitors	RSC Advances	2018	ISSN 2046-2069
Simpkins, Scott W. et al.	Predicting bioprocess targets of chemical compounds through integration of chemical-genetic and genetic interactions	PLoS Computational Biology	2018	ISSN 1553-7358
Nixon, Gemma L. et al.	Repurposing and reformulation of the antiparasitic agent flubendazole for treatment of cryptococcal meningoencephalitis, a neglected fungal disease	Antimicrobial Agents and Chemotherapy	2018	ISSN 1098-6596
Sato-Kaneko, Fumi et al.	Discovery of a Novel Microtubule Targeting Agent as an Adjuvant for Cancer Immunotherapy	BioMed Research International	2018	ISSN 2314-6141
Dilworth, David et al.	The prolyl isomerase FKBP25 regulates microtubule polymerization impacting cell cycle progression and genomic stability	Nucleic Acids Research	2018	ISSN 1362-4962
Huan, L C et al.	Exploration of Some Thiazolidine-2, 4-dione and 2-Oxoindoline Derivatives Incorporating 3, 4, 5-Trimethoxybenzyl Moiety as Novel Anticancer Agents	Letters in Drug …	2018	Article Link
Qi, Jianguo et al.	Synthesis and biological evaluation of N-substituted 3-oxo-1,2,3,4-tetrahydro-quinoxaline-6-carboxylic acid derivatives as tubulin polymerization inhibitors	European Journal of Medicinal Chemistry	2018	ISSN 1768-3254
Senese, Silvia et al.	Microtubins: A novel class of small synthetic microtubule targeting drugs that inhibit cancer cell proliferation	Oncotarget	2017	ISSN 1949-2553
Wieczorek, Anna et al.	Synthesis and evaluation of biological properties of ferrocenyl-podophyllotoxin conjugates	Dalton Transactions	2017	ISSN 1477-9234
Sun, Maolin et al.	Synthesis and bioevaluation of N,4-diaryl-1,3-thiazole-2-amines as tubulin inhibitors with potent antiproliferative activity	PLoS ONE	2017	ISSN 1932-6203
Zollo, Massimo et al.	PRUNE is crucial for normal brain development and mutated in microcephaly with neurodevelopmental impairment	Brain	2017	ISSN 1460-2156
Chen, Minfeng et al.	Pericyte-targeting prodrug overcomes tumor resistance to vascular disrupting agents	Journal of Clinical Investigation	2017	ISSN 1558-8238
Tantak, Mukund P. et al.	Design and synthesis of bis(indolyl)ketohydrazide-hydrazones: Identification of potent and selective novel tubulin inhibitors	European Journal of Medicinal Chemistry	2017	ISSN 1768-3254
Xu, Qile et al.	Design, synthesis and structure-Activity relationship of 3,6-diaryl-7H-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazines as novel tubulin inhibitors	Scientific Reports	2017	ISSN 2045-2322
Kowalczyk, Karolina et al.	Colchicine metallocenyl bioconjugates showing high antiproliferative activities against cancer cell lines	Dalton Transactions	2017	ISSN 1477-9234
Wu, Yue et al.	Design and synthesis of 5-aryl-4-(4-arylpiperazine-1-carbonyl)-2H-1,2,3-triazole derivatives as colchicine binding site inhibitors	Scientific Reports	2017	ISSN 2045-2322
Fu, Ying et al.	The contrasting catalytic efficiency and cancer cell antiproliferative activity of stereoselective organoruthenium transfer hydrogenation catalysts	Dalton Transactions	2016	ISSN 1477-9234
Cai, De et al.	YSL-12, a novel microtubule-destabilizing agent, exerts potent anti-tumor activity against colon cancer in vitro and in vivo	Cancer Chemotherapy and Pharmacology	2016	ISSN 1432-0843
Magalhaes, Luma G. et al.	Discovery of a series of acridinones as mechanism-based tubulin assembly inhibitors with anticancer activity	PLoS ONE	2016	ISSN 1932-6203
Hasanpourghadi, Mohadeseh et al.	Targeting of tubulin polymerization and induction of mitotic blockage by Methyl 2-(5-fluoro-2-hydroxyphenyl)-1H-benzo[d]imidazole-5-carboxylate (MBIC) in human cervical cancer HeLa cell	Journal of Experimental and Clinical Cancer Research	2016	ISSN 1756-9966
Pearson, Brandon L. et al.	Identification of chemicals that mimic transcriptional changes associated with autism, brain aging and neurodegeneration	Nature Communications	2016	ISSN 2041-1723
Yoshitake, Jun et al.	Modification of tau by 8-nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP): Effects of nitric oxide-linked chemical modification on tau aggregation	Journal of Biological Chemistry	2016	ISSN 1083-351X
Thapa, Pritam et al.	Far-Red Light-Activatable Prodrug of Paclitaxel for the Combined Effects of Photodynamic Therapy and Site-Specific Paclitaxel Chemotherapy	Journal of Medicinal Chemistry	2016	ISSN 1520-4804
Fukuda, Yasunori et al.	Tubulin is a molecular target of the Wnt-activating chemical probe	BMC Biochemistry	2016	ISSN 1471-2091
Kamal, Ahmed et al.	Design, synthesis and antiproliferative activity of the new conjugates of E7010 and resveratrol as tubulin polymerization inhibitors	Organic and Biomolecular Chemistry	2016	ISSN 1477-0520
Fawzy, Iten M. et al.	Newly Designed and Synthesized Curcumin Analogs with in vitro Cytotoxicity and Tubulin Polymerization Activity	Chemical biology & drug design	2015	ISSN 1747--0285
Mu, Yan et al.	The novel tubulin polymerization inhibitor MHPT exhibits selective anti-tumor activity against rhabdomyosarcoma in vitro and in vivo	PLoS ONE	2015	ISSN 1932-6203
Xu, Qile et al.	Synthesis and biological evaluation of 3-alkyl-1,5-diaryl-1H-pyrazoles as rigid analogues of combretastatin A-4 with potent antiproliferative activity	PLoS ONE	2015	ISSN 1932-6203
Seashore-Ludlow, Brinton et al.	Harnessing connectivity in a large-scale small-molecule sensitivity dataset	Cancer Discovery	2015	ISSN 2159-8290
Remers, William A. et al.	Synthesis and Antitumor Activity of Heterocycles Related to Carbendazim	Journal of Heterocyclic Chemistry	2015	ISSN 1943--5193
Sheldon, Jonathon E. et al.	Photoswitchable anticancer activity via trans-cis isomerization of a combretastatin A-4 analog	Organic and Biomolecular Chemistry	2015	ISSN 1477-0520
Calles, Antonio et al.	Tivantinib (ARQ 197) efficacy is independent of MET inhibition in non-small-cell lung cancer cell lines	Molecular Oncology	2015	ISSN 1878-0261
Yan, Jun et al.	A novel synthetic compound exerts effective anti-tumour activity in vivo via the inhibition of tubulin polymerisation in A549 cells	Biochemical Pharmacology	2015	ISSN 1873-2968
Wu, Shaoyu et al.	Bis-cyclopropane analog of disorazole C1 is a microtubuledestabilizing agent active in ABCB1-overexpressing human colon cancer cells	Oncotarget	2015	ISSN 1949-2553
Shigehiro, Tsukasa et al.	Efficient Drug Delivery of Paclitaxel Glycoside: A Novel Solubility Gradient Encapsulation into Liposomes Coupled with Immunoliposomes Preparation	PLOS ONE	2014	ISSN 1932--6203
Choi, Bo Hwa et al.	Suprafenacine, an Indazole-Hydrazide Agent, Targets Cancer Cells Through Microtubule Destabilization	PLOS ONE	2014	ISSN 1932--6203
Senese, S. et al.	Chemical dissection of the cell cycle: Probes for cell biology and anti-cancer drug development	Cell Death and Disease	2014	ISSN 2041-4889
Lutz, Vanessa et al.	SAR studies on hydropentalene derivatives - Important core units of biologically active tetramic acid macrolactams and ptychanolides	Bioorganic and Medicinal Chemistry	2014	ISSN 1464-3391
Mei, Mei et al.	A new 2α,5α,10β,14β-tetraacetoxy-4(20),11-taxadiene (SIA) derivative overcomes paclitaxel resistance by inhibiting MAPK signaling and increasing paclitaxel accumulation in breast cancer cells	PLoS ONE	2014	ISSN 1932-6203
Bio, Moses et al.	Far-red light activatable, multifunctional prodrug for fluorescence optical imaging and combinational treatment	Journal of Medicinal Chemistry	2014	ISSN 1520-4804
Mäki-Jouppila, Jenni H.E. et al.	Centmitor-1, a novel acridinyl-acetohydrazide, possesses similar molecular interaction field and antimitotic cellular phenotype as rigosertib, on 01910.Na	Molecular Cancer Therapeutics	2014	ISSN 1538-8514
Zhang, Qiu et al.	Anti-tumor selectivity of a novel Tubulin and HSP90 dual-targeting inhibitor in non-small cell lung cancer models	Biochemical Pharmacology	2013	ISSN 1873-2968
Bio, Moses et al.	Site-specific and far-red-light-activatable prodrug of combretastatin A-4 using photo-unclick chemistry	Journal of Medicinal Chemistry	2013	ISSN 0022-2623
Zach, Frank et al.	The retinitis pigmentosa 28 protein FAM161A is a novel ciliary protein involved in intermolecular protein interaction and microtubule association	Human molecular genetics	2012	ISSN 1460--2083
Sidhaye, Venkataramana K. et al.	A Novel Role for Aquaporin-5 in Enhancing Microtubule Organization and Stability	PLOS ONE	2012	ISSN 1932--6203
Nguyen, Tam Luong et al.	Evading Pgp activity in drug-resistant cancer cells: A structural and functional study of antitubulin furan metotica compounds	Molecular Cancer Therapeutics	2012	ISSN 1535-7163
Fitzgerald, Daniel P. et al.	TPI-287, a new taxane family member, reduces the brain metastatic colonization of breast cancer cells	Molecular Cancer Therapeutics	2012	ISSN 1535-7163
Kawaratani, Yasuyuki et al.	New microtubule polymerization inhibitors comprising a nitrooxymethylphenyl group	Bioorganic and Medicinal Chemistry	2011	ISSN 0968-0896
Dyrager, Christine et al.	Inhibitors and promoters of tubulin polymerization: synthesis and biological evaluation of chalcones and related dienones as potential anticancer agents	Bioorganic & medicinal chemistry	2011	ISSN 1464--3391
Hwang, Ji Hwan et al.	Induction of tubulin polymerization and apoptosis in malignant mesothelioma cells by a new compound JBIR-23	Cancer letters	2011	ISSN 1872--7980
Kim, Insook et al.	Zinc stimulates tau S214 phosphorylation by the activation of Raf/mitogen-activated protein kinase-kinase/extracellular signal-regulated kinase pathway	NeuroReport	2011	ISSN 0959-4965
Oussenko, Irina A. et al.	Effect of on 01910.Na, an anticancer mitotic inhibitor, on cell-cycle progression correlates with RanGAP1 hyperphosphorylation	Cancer Research	2011	ISSN 0008-5472

Faqs

Question 1: What are the advantages to using this kit?

Answer 1: BK011P is a fluorescence-based tubulin polymerization assay kit. Compared to the absorbance-based kits, BK011P has increased sensitivity, signal-to-noise ratio and an improved coefficient of variation. The greater sensitivity allows the researcher to use 1/3 as much tubulin which means that the BK011P kit provides 96 assays versus the 30 assays of BK004P or BK006P. On a cost per assay basis, BK011P is the best value of the tubulin polymerization kits.

Question 2: Which kit is best for screening a compound for it’s effects on tubulin polymerization?

Answer 2: All 3 tubulin polymerization kits (2 absorbance-based kits, BK004P and BK006P; 1 fluorescence-based kit, BK011P) are well-suited for screening of potential tubulin polymerization enhancers and inhibitors. Each kit has its own pros and cons. For initial compound/drug screening, we recommend the absorbance-based tubulin polymerization assay BK004P which is the most economical. This kit uses 97% pure tubulin (remaining 3% are MAPs) while BK006P and BK011P use >99% pure tubulin. This is an important difference because the presence of MAPs means that tubulin polymerization can be examined in the absence of enhancers or inhibitors with as little as 3 or 4 mg/ml tubulin using the BK004P kit. To study enhancers, we recommend using 3 mg/ml tubulin, whereas 4 mg/ml tubulin is recommended for inhibitors. In the case of BK004P, MAPs act as polymerization enhancers. With BK006P and BK011P, an enhancer such as glycerol or taxol must be used to drive tubulin polymerization with concentrations <5 mg/ml tubulin. Using tubulin at 5 mg/ml or higher allows for the omission of glycerol or taxol, but requires additional tubulin. In some cases, glycerol can interfere with the binding of tubulin accessory proteins or compounds. However, since BK011P is fluorescence-based, there is increased sensitivity that allows the researcher to use 1/3 as much tubulin with greater sensitivity. Thus, the kit provides 96 assays versus the 30 assays of BK004P or BK006P. Assay conditions can easily be altered to test for glycerol interference.

If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com

BE THE FIRST TO KNOW

Tubulin polymerization assay using >99% pure tubulin, fluorescence based (BK011P)

Events, Sales, And Offers