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What can Silensomes™ HLM be used for?

Silensomes™ HLM can be used in cytochrome P450 screening and phenotyping assays, to identify enzymes responsible for the metabolism of drug candidates. Silensomes™ HLM can be used to predict directly the in-vitro fm for a particular CYP450 enzyme (fm...

Which program should I use for co-transfection of RNP and CRISPR/Cas9 complexes ? The paper "Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells from Aikiko Seki and Sascha Rutz showed different programs as you have in the Lonza optimized Nucleofector Protocols?

Their publication in the Journal of Experimental Medicine is an advanced optimization for gene knock out in primary human and mouse T cells. The final protocol achieved 85-98% gene knock out for several tested receptors.The group has tested and...

What is CYP450 phenotyping?

In vitro identification and measurement of the contribution of the major cytochrome P450 enzymes (CYP450s) involved in the metabolism of a new drug candidate is called “CYP450 phenotyping”. Information from these assays is used to predict the impact...

Can I modify the RAFT™ Standard Protocol?

Yes. The standard protocol is an excellent starting point for using the RAFT™ System however the system allows the researcher to modify the conditions to better suit their cells and experimental design. For instance the volume of collagen or the...

Which Nucleofector™ Protocol should I use for genome editing and transfection of CRISPR/Cas9 into my cells?

Please use always the ready-to-use protocol for the specific cell type. The optimized programs are cell type specific and do work for any kind of substrates. You can transfect small and large DNA vectors, mRNA, siRNA, gRNA, peptides and proteins with...

What are "Control Silensomes™ HLM"?

Control Silensomes™ HLM are the true control microsomes for comparison of metabolism in “CYP-silenced” Silensomes™ HLM since they have been processed under the same conditions as Silensomes™ HLM but in the absence of the mechanism based inhibitor...

How can I image cells in a RAFT™ 3D Cell Culture?

Below is a list of compatible imaging techniques and systems with RAFT™ Cultures, depending on the type of cells being cultured. RAFT™ Culture Types: Cells that tend to maintain a round morphology or that form higher order structures such as...

What is fm value and why is important to have that data in-vitro/in-vivo?

The fm value is the percent of enzyme (e.g. CYP) contribution to the metabolism of a test compound. The in-vitro fm value is important to measure to predict the in vivo fm which is important to estimate the potential for drug-drug interactions with...

Can I remove the RAFT™ 3D Cell Cultures from the plate in which they have been made?

This is easily done using a pair of forceps. First, pass the forceps around the edge of the culture to help dislodge it from the bottom of the well, and then pinch the edge of the culture lightly to avoid damaging the culture. When in contact with...

How can in-vitro fm value be estimated using Silensomes™ HLM?

The Silensomes™ HLM directly predict the CYP contribution to drug metabolism and the in-vitro fm. .This is carried out by measuring the intrinsic clearance (Clint) of a test compound in Silensomes™ and in concurrent Control SilensomesTM HLM. The...

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