Cell Biology

Magnetofection™ for Enhanced Transfection Efficiency in Primary and Hard-to-Transfect Cells

Magnetic Assisted Transfection is a novel technology developed by OZ Biosciences. It has a proven high efficiency in delivering molecules into challenging cells. 
Magnetic nanoparticles are associated with nucleic acids (naked or pre-complexed with a transfectionreagent or viruses) by salt-induced aggregation and electrostatic interactions.
Magnetic force drives these complexes towards the target cells, allowing a rapid concentration of the vector dose onto cells
The cellular uptake of the genetic materials is accomplished by endocytosis and pinocytosis
Nucleic acids are released in the cytoplasm by flip-flop mechanism or proton sponge effect*

Magnetofection data

 

* Plank et al , Adv. Drug Deliv. Rev. (2011), 63(14-15):1300-31

 

New Transfection and Transduction Reagents with Unique Technologies

 

The reagents from OZ Biosciences covers a large range of applications:
Transfection Reagents
Viral Applications
Genome Editing
Protein Delivery
In vivo Delivery
Cellular Assays
Vaccine Adjuvants
Transfection Tools

Get catalogue

 

Laminar Wash Sample Preparation for Single-Cell Sequencing

Single-cell genomics is rapidly growing. Nature Methods named multimodal omics the method of the year for 2019. And as more scientists use single-cell techniques, they find that sample preparation is very important to achieve high-quality data.

Single cell sequencing can suffer from poor data quality due to clumping of cells, incomplete washing of cells leaving cell-free nucleic acids, and poor retention of low numbers of cells. Washing via centrifugation causes stress on the cells in samples and can cause erroneous gene expression results.

Sample prep is extremely important in single-cell genomics, especially for single cell gene expression and immune profiling. In this area, mRNA transcripts are being barcoded and cells can experience mRNA leakage (causing increased background) or mRNA degradation (causing less signal). Unlike with flow cytometry, in single-cell sequencing it is not possible to gate out debris or groups of cells. The cell preparation must be as clean as possible and consist of single cells.

By using the Laminar Wash system scientists can see time savings and less manual manipulation.

Laminar Wash systems can wash cells without putting stress on cells that may impact gene expression, cell integrity and viability.

Laminar Wash™ Technology Overview

 

Nuclei Isolation Using Laminar Wash

PBMCs were lysed in bulk in tube with Lysis Buffer, quenched with Wash Buffer and centrifuged
Cells are further either washed twiced in tube by centrifuge, OR plated in Laminar Wash plate to settle for 20min, washed 7x in a Laminar Wash HT1000 System with PBS + 0.04% BSA

CAR-T-Cell Production

Reliable, Automated and Reproducible Flow Cytometry

The unique Laminar Wash™ technology simplifies and automates flow cytometry workflows for CAR-T-cell manufacturing. Data improves reproducibility, saves time and gives tighter populations. Laminar Wash™ replaces centrifugation-based stain&wash and is more gentle to cells. Time saving in this workflow was 50 min/assay.

CAR-T cell therapy biotech company AdiCet Bio improved data by automating of a part of their flow cytometry assay. They saw improved consistency and are planning full automation using a Laminar Wash™ system.

Download AdiCet Bio Case Study

Laminar Wash™ Technology Overview

 

Change the way you process cells to get reliably consistent data in flow cytometry

The DA-Cell Laminar wash technology replaces centrifugation by using a novel DA-Cell technology, which is gentle to cells, giving higher retention.

  • A wide range of cell numbers, from single-cell to 10 million cells per well
  • No stress on cells from centrifugation and improved viability
  • Reduced debris and aggregation of cells
  • Cleaner and faster washing

DA-Cell technology page

Enhanced viability and retention

Cells from DA Cell method demonstrate much stronger fluorescence signal and intact morphology of cells

Centrifugation presumably causes  significant stress on cells distorting its physiology and protein expression

This is how it works!

Check our video describing the efficient laminar wash of suspension cells for flow cytometry

VIROMER® Cytostain Organelle-Targeted Live Cell Imaging

Fluorescence staining of selected organelle
Down-stream experiments unaffected – non-toxic and biological
Easy and straightforward

The new Viromer Cytostain adds content at great convenience at low toxicity. With the mRNA encoding fluorescent fused protein expression, you will stain your targeted organelle of interest in living cells without impacting viability. Your flexibility for downstream experiments are therefore retained.

 

 

 

 

Viromer Cytostain – how does it work?

The Viromer Cytostain reagent is a Viromer nanoparticle, complexed to mRNAs encoding for fluorescent proteins. Thanks to the features of the Viromer particle, the mRNA is efficiently delivered into the cell cytosol and the protein is directly expressed.

Viromer is an efficient, non-toxic polymer which is used for transfection of both standard and hard-to-transfect cells. The team at Lipocalyx has extensive experience in the development of tools for cell transfection, including the transfection of mRNA.

Protocol Viromer Cytostain

Straightforward protocol; rehydrate and add to cells. Signal detachable in 6 hours.

 

Cytostain product page

Next Generation Viromer® Transfection
Viromer® mRNA

The new Viromer formulation has developed for higher efficiencies and easier protocols.
It is a versatile tool for non-toxic transfection of mRNA into a broad range of standard and challenging cells. mRNA transfection has several benefits, in particular when plasmid DNA delivery and uptake is inefficient due to natural defence mechanisms against foreign dsDNA, such as immune cells.

Speed: mRNA does not need to reach the nucleus for cellular action and translation occurs through a promoter-independent process and the desired pro­tein is detectable as early as 6 h post-transfection.

Safety: No risk of genomic integration and transient expression avoids toxicity related to protein accumulation.

Productivity: As simple as in nature, mRNA instructs directly cells to produce proteins. With an optimized delivery, it is then possible to reach very high expression levels of any intracellular, transmembrane or excreted protein.

Place your order today!

 

Next Generation Viromer® Transfection
Viromer® Plasmid

The new Viromer formulation has developed for higher efficiencies and easier protocols.
It is a versatile tool for non-toxic transfection of pDNA into a broad range of standard and challenging cells.
Viromer Plasmid serves as a great option to electroporation.

Place your order today!

 

VIROMER® CRISPR for RNP delivery

Genome-editing using Cas9/gRNA ribonucleoprotein

Pre-formed RNP complex enables a direct action of the CRISPR system upon delivery. There is no need to pass through the transcription/translation cell machinery for expressing the Cas9 protein, and it skips the assembly step with the gRNA into the cytosol. It is then faster and safer!
Second, the cell rapidly clears Cas9/gRNA RNPs through protein degradation pathways.
This transient action provides a certain degree of control as it greatly increases the fidelity of the endonuclease (less off-target cleavage) and there is no risk of integration into the cell genome.

 

Download Viromer CRISPR LabBook
More data

 

 

 

siRNA Off-Target Effects; Causes, Extent & Impact
siPOOLs™ are innovative, complex siRNA pools which reduce false positives & false negatives in siRNA knockdown experiments

Screen Shot 2017-10-03 at 16.30.27

Individual siRNAs or low complexity siRNA pools containing 3-4 siRNAs often hit multiple off-target genes and exhibit variable target gene knock-down. Short interfering RNA (siRNA) pools, siPOOLs are highly complex and defined pools of 30 siRNAs, each siRNA present at picomolar working concentrations.

Benefits:
dilutes the off-target signature of each siRNA, increasing on-target specificity
ensures co-operative knock-down of the target gene, producing more robust and reliable results.

Download the new TechNote:
Off-Target Effects; Causes, Extent & Impact

Publication:
Hannus et al NAR

 

Screen Shot 2017-10-03 at 16.36.23

Screen Shot 2017-10-03 at 16.36.07

 

Get Higher Protein Expression by Transfecting mRNA instead of pDNA in Macrophages and Monocytes

Transfecting cells with mRNA greatly improves protein expression levels in immune cells that have innate defence mechanisms against foreign DNA. Being a natural component of the cytosol, mRNA will stay intact and create a productive translation process. Viromer® RED and Viromer® YELLOW transfection reagents are designed to work well with both mRNA and pDNA. No special, expensive transfection reagent is required!

Screen Shot 2017-05-09 at 16.26.44

Viromer® Start Positive® Controls helps you to compare mRNA and pDNA transfection!
Pre-formulated transfection complexes for use with either Viromer® RED or Viromer® YELLOW
Includes both a GFP-encoding mRNA- and a pCMV-GFP plasmid transfection complex
A perfect tool to evaluate the optimal conditions for your cells!

Comparison of mRNA and pDNA transfection, using Start Positive Controls:

Screen Shot 2017-05-09 at 16.29.22
Ordering information:
The Start Positive Controls are available in two formats based on the different transfection reagents Viromer RED and Viromer YELLOW. Both include the pDNA-GFP plasmid, as well as GFP-encoding mRNA.
Viromer RED is versatile for standard and challenging cells, whilst Viromer YELLOW can be optimal for specific cells. Please contact us if you are unsure which is best for you.

VR-BUNDLE-01, Viromer RED pDNA/mRNA-GFP controls
VY-BUNDLE-01, Viromer YELLOW pDNA/mRNA-GFP controls
Price: SEK 683.-; DKK 525.-; NOK 683.-; € 70.-
Place your order here: mailto@lablifenordic.com
Screen Shot 2017-05-09 at 16.29.47

Want to learn more about Viromer® Transfection Reagents from Lipocalyx?
Get a copy of the FactBook here:Viromer-transfection-factbook

Lipocalyx_blau

Maximise Your Transfection Efficiency with Viromer®

Optimising transfections in four steps. With Viromer reagents, delivery of the complex is effective and cells are treated gently.

In our new LabBook, you get access to recommendations for both adherent and suspension cells.
Optimisation guidelines are based on using the following four main parameters:
Amount of complex
Cell density & confluency
Viromer – RNA/DNA ratio
Incubation time

Get you copy of the LabBook here:
pDNA/mRNA: LabBook-Viromer-RED-YELLOW
siRNA/miRNA: LabBook-Viromer-BLUE GREEN

Screen Shot 2017-03-22 at 19.40.56 Screen Shot 2017-03-22 at 19.40.43

 

This is how the Viromer technology works:

Article: GEN#022 volume 34 #4 pg 20-21

Use Viromer RED and Viromer YELLOW for pDNA and mRNA, Viromer BLUE and Viromer GREEN for siRNA and miRNA.
Order your Viromer transfection reagent here: mailto@lablifenordic.com

 

Viromer® One Red; Evolution in mRNA and Plasmid Transfection

Instead of manual preparations, Viromer® ONE RED is an easy, individual and reliable new transfection reagent which brings industry standard into your lab.

  • Viromer® ONE RED is fully compatible with our Viromer® RED liquid product, but is easier to handle and significantly reduces variability. Please refer to our cell data base for a complete list of cell types that are amenable to Viromer transfection.
  • ONE Step Transfection.
  • High transfection efficiency due to an active escape of Viromer® complexes from the endosome.
  • Great safety because Viromer complexes are non-charged, gentle on cells and compatible with serum and antibiotics.
  • Easy and fast transfection with consistent results ascribed to a one step protocol including initial optimization.

 

Comparative data:
Any cell. Any cargo. Viromer® ONE RED is a highly potent and universal reagent for the transfection of both DNA or mRNA.

Relative transfection efficacy. The highest efficacy on each cell line was set to 100%. Standard conditions according to suppliers protocol (n=8).

viromer-transfection-one-competes

Protocol here: transfection-protocol-Viromer-ONE-RED

Publications for mRNA/Plasmid Transfection with Viromer Red

Defining functional interactions during biogenesis of epithelial junctions
Erasmus et.al., Nat Commun. 2016
Extensive nuclear sphere generation in the human Alzheimer’s brain
Kolbe et. al., NeuroBiolAging, 2016
Hepatic Fgf21 Expression Is Repressed after Simvastatin Treatment in Mice.
Ziros et. al., PLoS One., 2016
Systemic RNA delivery to dendritic cells exploits antiviral defence for cancer immunotherapy.
Kranz et. al., Nature. 2016
Delta-Like Ligand 4 Modulates Liver Damage by Down-Regulating Chemokine Expression.
Shen et. al., Am J Pathol., 2016
Tumor suppressor BTG1 promotes PRMT1-mediated ATF4 function in response to cellular stress.
Yuniati et. al., Oncotarget, 2016
Drug Delivery Using Novel Biological and Synthetic Materials
Ito et. al.Biomed Res Int, 2015
The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells
Rao et.al., Hindawi Publishing Group, BioMed Research International, 2015

 

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