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Can you retrieve the cells from the RAFT Collagen Matrix by dissociation?

It is possible to dissociate the cells from the collagen matrix, by treating them with collagenase I or IV, both of which cleave the bond between a neutral amino acid (X) and glycine in the sequence Pro-X-Gly-Pro, which is found with high frequency...

What are the parameters that affect the accuracy of my fm value?

The parameters that affect the accuracy of in-vitro fm value calculation include: 1) the choice of analytics (e.g. disappearance of parent vs appearance of metabolite), 2) the activities in the initial HLM batch, and 3) the balance between the CYPs....

Can I change the concentration of collagen? How thick or thin can the RAFT System get the collagen layer?

The standard RAFT™ Protocol yields a culture of 100-120µm thickness at a collagen concentration of about 80mg/mL. It is possible to increase the collagen concentration to 100mg/mL by leaving the RAFT™ absorber in contact with the hydrogel for 30...

What is the regulatory position of the fm value?

The FDA and EMA guidelines consider metabolism to be a significant pathway when it contributes to 25% or more of the drug’s overall elimination. When the CYP contribution is ~25% (In-vitro fm value > 25 %) or unknown, in vivo studies using...

Do you have a Nucleofection Protocol for bacterial transformation for the Nucleofector I, IIN, IIS or 2b Device?

Yes, we do have guidelines for bacteria transformation on the Nucleofector™ Device. However, not all models of the Nucleofector™ Device series are capable of bacteria transformation and have bacteria pulses implemented in the program list, only the...

What are the Storage and thaw-refreeze conditions for Silensomes HLM?

The Silensomes™ HLM and associated Control Silensomes™ HLM should be stored at or below -70°C. CYP450 enzyme activities in Silensomes™ HLM and associated Control Silensomes™ HLM are stable for up to 5 years when stored at -70°C and can be...

How can we approach the in-vitro fm value for low clearance compounds when using Silensomes HLM?

The in-vitro fm value of low clearance compounds can be measured by optimizing or extending the incubation time with certain limit. In addition, the appearance of metabolites can be measured instead of the disappearance of the parent compound.

What is the benefit of using Silensomes instead of the classical testing methods?

Compared to classical phenotyping protocols relying on reversible, as well irreversible, inhibitors Silensomes™ HLM offer a superior performance and high level of quality. The mechanism based inhibitors used to make Silensomes™ HLM are irreversible,...

Do you have a Nucleofector protocol for transfection of yeast (Saccharomyces cerevisiae)?

Unfortunately we do not have any records about Nucleofection of yeast. It is also impossible to search for current publications, because you get only false positive hits. A lot of yeast genes have been nucleofected and many reportergene systems have...

How reproducible is the fm data for Silensomes HLM?

The fm value consistency depends on consistency in the analytical method used to calculate the disappearance or appearance. If the disappearance of the test compound is lower than 5 or 10% over the incubation time such as with low clearance compound,...
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