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Question 1: What is the difference between 70% pure porcine brain tubulin ((Cat. #ML116) and 70% pure bovine brain tubulin (Cat. # ML113)?
Answer 1: The only difference is the source of the tubulin: bovine vs porcine brains. Functionally, there are no significant differences between tubulin isolated from bovine brains vs porcine brains. Stringent quality control and in-house testing has revealed no differences in polymerization, response to drugs (enhancers and inhibitors) or binding to accessory proteins (e.g., motor proteins). (See Comparison of Porcine and Bovine tubulin)
Question 2: Does tubulin polymerization using 70% pure porcine brain tubulin/30% MAPs ((Cat. #ML116) require additional enhancers such as glycerol or taxol?
Answer 2: No additional enhancers are required when using 2 mg/ml of the MAP-enriched 70% pure tubulin. The recommended polymerization reaction using 70% pure tubulin contains 180 &mu, l of 2 mg/ml tubulin in 80 mM PIPES pH 6.9, 0.5 mM EGTA, 2 mM MgCl2 and 1 mM GTP. Polymerization is started by incubation at 37°C and followed by absorption readings at 340 nm. Under these conditions polymerization reaches a maximal OD340 between 0.3 - 0.4 within 20 minutes. In this experimental set up (180 &mu, l volume in a spectrophotometer with a pathlength of 0.8 cm using Corning Costar&rsquo, s half area well plate Cat. # 3696) an OD340 of 0.16 is approximately equal to 1 mg per ml of tubulin polymer mass. Thus, under the conditions described, approximately 100% of the tubulin is polymerized. If polymerization enhancers such as taxol are to be used or tested, we recommend reducing the tubulin concentration to 1 mg/ml. These conditions will result in a more pronounced nucleation phase and a lower polymerization curve.
Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.
All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.
ML116-B.pdf
ML116-B-datasheet.pdf
ML116-B-safety-datasheet.pdf
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