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Tubulin protein (95% pure): porcine brain

Tubulin protein (95% pure): porcine brain

*Amount

1x4 mg (Cat. # HTS03-A) +$250.00
1x40 mg (Cat. # HTS03-B) +$1,350.00
1x100 mg (Cat. # HTS03-XL) +$2,650.00

Price: $0.00

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Product Uses Include

Economical alternative to T240 in primary screens for tubulin ligand drugs

High-throughput tubulin polymerization or depolymerization screens

Production of microtubule substrates for HTS motor assays

Material
Tubulin protein has been isolated from porcine brain. The final product contains approximately 97% tubulin and 3% Microtubule Associated Proteins (MAPs). This version of porcine tubulin is an economical alternative to our highly purified tubulin (Cat. # T240) for anti-tubulin ligand drug discovery. It has been formulated to be compatible with 96 or 384-well format polymerization assays such as in Biochem™ kits BK004P or BK011P .

Purity
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 12% SDS-PAGE gel. HTS02 contains 97% tubulin (MW. 55 kDa) and 3% MAPs (MW 35-280 kDa).


	Figure 1: A 100 µg sample of HTS03 protein was separated by electrophoresis on a 12% SDS-PAGE gel and stained with Coomassie Blue. Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat.# ADV02).

Biological Activity
An ASSAY UNIT is defined as the amount of HTS03 protein needed to achieve a tubulin polymerization signal of OD340 of 0.1 - 0.15 in 30 minutes at 37°C in G-PEM buffer. The assay volume is 100 µl and assumes a spectrophotometer pathlength of 0.5 cm. The protein concentration for HTS02 in this assay is approximately 4 mg/ml. The biological activity of HTS03 is assessed by a tubulin polymerization assay. One ASSAY UNIT of tubulin is used for each polymeriztion assay. An OD340 of 0.10 - 0.15 is required to pass quality control. Tubulin polymerization must also be responsive to polymerization enhancers (paclitaxel) and inhibitors (nocodazole) at 5 µM drug concentration.


	Figure 2: Tubulin polymerization in a 96-well format using HTS03. All samples are in duplicates. Each well has a mini-polymerization curve with 20 min on the x-axis and 0.30 OD 340 nm on the y-axis. The V_max parameter is used to compare inhibitors, the CV is 13% in this format, so a 50% cut off is required for a 98% confidence.

Tubulin protein (95% pure): porcine brain

Tubulin protein (95% pure): porcine brain

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