Tubulin protein: MCF-7 cancer cell

Tubulin protein: MCF-7 cancer cell
$0.00

* Limited stock available.  If stock is not available, Cytoskeleton will produce a new batch upon request.  Minimum order will apply.  Inquire for more information.

 

Product uses

  • Screening for anti-tumor drugs
  • Assessing the effects of modulators of microtubule dynamics on tubulin isolated from actively growing cells
  • A new substrate for motor proteins

Material
MCF-7 cell tubulin is isolated from the human breast cancer cell line MCF-7 (ATCC, cat. # HTB-22), a model system to study many aspects of tumor cell growth. MCF-7 cell tubulin may be used in all situations where previously bovine brain tubulin has been employed, for example drug screening, motility assays and biochemical studies including microtubule dynamics. The advantage of using this novel tubulin is that it is derived from an actively dividing cell line, which often, in contrast to brain derived tubulins, more accurately portrays the situation that many researchers are trying to reconstruct in vitro.

The specificity of ligands for for a particular tubulin variant can be determined by performing comparative studies with both cancer cell and neuronal tubulins. Cytoskeleton, Inc. has advanced this concept by developing the Tubulin Ligand Index (TLI) system (patent pending). In this system, IC50 values for inhibitory compounds or EC50 values for stabilizing compounds are determined in in polymerization assays using cancer cell and neuronal tubulins. The IC50 or EC50 values for each tubulin variant are analyzed as a ratio (neuronal/cancer cell) and allow for determinations of the relative specificity for each tested compound. TLI values greater than 1.0 indicate that the particular compound is more active on cancer cell tubulin. Conversely, TLI values less than 1.0 suggest that a compound is more specific for neuronal tubulin. Table 1 summarizes data from a study comparing the specificity of several tubulin ligands using the TLI system.

Table 1. Tubulin ligand index values from studies with cancer cell and neuronal tubulins

EC50/IC50 Value (µM)
TLI Value
Ligand
Neuronal
MCF-7
HeLa
MCF-7
HeLa
Paclitaxel
Docetaxel
10-Deacetyl Taxol
0.48
0.47
3.71
0.51
0.34
4.20
1.04
0.41
30.00
0.94
1.38
0.88
0.46
1.15
0.12
Vinblastine
Vincristine
1.10
1.58
1.21
n.d.
2.83
2.25
0.91
n.a.
0.39
0.70
Colchicine
Nocodazole
Mebendazole
MF708
4.10
3.40
3.98
3.54
4.60
3.20
14.80
n.d.
3.10
3.20
25.00
1.91
0.89
1.06
0.27
n.a.
1.32
1.06
0.16
1.85

The protein is supplied as a lyophilized powder.

For another human cancer cell derived tubulin, see our HeLa cell tubulin (Cat. # H001)

Purity
H005 contains >90% pure MCF-7 cell tubulin. Purity is determined by scanning densitometry of proteins on SDS-PAGE gels.


Figure 1: An H001 sample (20 µg) was run on a 10% SDS-PAGE gel and stained with Coomassie Blue stain

Biological Activity
One unit of tubulin is defined as 24 µg of purified protein (as determined by the Precision Red Advanced Protein Assay Reagent, Cat. # ADV02). MCF-7 cell tubulin will polymerize efficiently at a concentration of 2.0 mg/ml in the presence of 20% (v/v) glycerol. The MCF-7 tubulin polymerization assay has been miniaturized to 12 µl (1 unit/reaction) in order to make anti-cancer drug development feasible. Visit the page on CytoDYNAMIX screens for more information. Tubulin from cervical cancer cells (HeLa) is also available (Cat. # H001).

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

M.S. Feizabadi and Y. Jun. 2014.Kinesin-1 translocation: Surprising differences between bovine brain and MCF7-derived microtubules. Biochem. Biophys. Res.Commun. DOI: 10.1016/j.bbrc.2014.10.119.

M.L. Antony et al. 2014. Growth arrest by the antitumor steroidal lactone withaferin A in human breast cancer cells is associated with downregulation and covalent binding at cysteine-303 of β-tubulin. J. Biol. Chem.

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