Highly efficient transfection of primary cells and cell lines
PromoFectin provides highly efficient and reproducible delivery of nucleic acids into a variety of cell types, including many hard-to-transfect cell lines and primary cells. It consists of a non-liposomal polymer that is stable, water-soluble and free of any components of animal origin. PromoFectin shows very low toxicity and is therefore ideally suited for the transfection of sensitive cells, yielding optimal transfection efficiencies and gene expression. Transfection with PromoFectin requires no medium change and it can be used for adherent and non-adherent cells. It compacts and protects the nucleic acid of interest, enables efficient transport into the cells via endocytosis and allows for a rapid and almost complete release of the intact nucleic acid into the cytosol – all of which favor the delivery to the nucleus.
Special cell type specific PromoFectin variants are also available, which have been developed for the efficient but gentle transfection of cells such as endothelial cells (e.g. primary HUVECs), hepatocytes, macrophages, or neuronal cells.
More products for cell transfection as well as gene cloning, expression and silencing as well as promotions can be found on our website.
Beckman Coulter Life Sciences Introduces a 561 nm Laser Option for New Gallios Flow Cytometers at CYTO 2012 - Range of Detected Dyes and Fluorescent Proteins Expands
A 561 nm laser option, available for new 561 nm-laser ready Gallios* flow cytometers from Beckman Coulter Life Sciences, will be on view at CYTO 2012, June 23-27 in booth 319 at the Congress Center in Leipzig, Germany. The 561 nm laser option provides researchers with the ability to analyze a greater range of fluorescent proteins and to more efficiently excite PE/PE tandems. Optimal excitation is offered for red fluorescent proteins such as mCherry and DsRed, which are being increasingly used in research labs.
Yellow-green excitation provided at 561 nm produces less autofluorescence in cells, increasing the signal-to-noise ratio for PE and DsRed detection. Since 561 nm excitation is well separated from fluorescein and GFP emission, there is virtually no crossover contamination in the green detection channel, allowing simultaneous fluorescein, or GFP detection, without blocking optics.
The new laser is externally coupled through an optical fiber and can be field installed on new Gallios 561 nm-laser ready analyzers. The 561 nm laser will be passed to the flow cell collinearly with the 488 nm laser during acquisition.
Available configurations for the Beckman Coulter Life Sciences’ Gallios flow cytometers are: 6 colour/2 laser, 8 colour/2 laser, 10 colour/3 laser and 10 colour/4 laser – the last configuration reflecting the 561 nm option. Six fluorescence detectors provide simultaneous acquisition of up to six fluorescence signals. Concurrent measurements of integral, peak and width are available for all parameters, including scatter and fluorescence signals.
“Expanding the colour options on the Gallios cytometer will extend the applications of the instrument for research scientists,” said Mario Koksch, director of project management and strategy at Beckman Coulter Life Sciences. “Researchers will be able to further refine their studies using the full capabilities of the cytometer.”
Gallios cytometers feature an innovative forward-scatter detector that provides up to three measurements of cell size and visualization of particles down to 0.404 mm in diameter. A side-scatter detector incorporates an independently focused, high-performance photodiode with electronic attenuation. A selection of up to 62 parameters can be processed per analysis, at acquisition rates of 25,000 events-per-second, with high yield. Easily interchanged optical filters facilitate detection of a variety of dyes and wavelengths.
Lonza introduces Nucleofector™ PLUS Supplements, a new add-on product for use in combination with Nucleofector™ Kits allowing Cryopreservation of Cells.
These specialized supplements which are used instead of the standard kit supplement allow cryopreservation of freshly isolated or harvested cells in Nucleofector™ Solution.
“By decoupling cell harvesting and transfection, Nucleofection™ Experiments can be done at the scientist’s convenience. Cells can be collected, stored and are ready-to-use at a later time, and deliver transfection results comparable to fresh cells. Larger cell batches can be cryopreserved so that researchers can avoid batch-to-batch variances in transfection results. Thus, Nucleofector™ PLUS Supplements can make whole experimental programs more consistent.” explains Dr. Herbert Mueller-Hartmann, Head of R&D Discovery Technologies.
Nucleofection™ is a transfection technology that was first introduced into the market in 2001. It is based on the unique combination of electrical parameters and cell-type specific solutions. It is known as the first efficient non-viral transfection method for primary neural cells and has been stated in over 300 peer-reviewed publications as the method of choice for transfection of primary neurons.
Designed for efficient, small-scale purification and screening of histidine-tagged proteins from different sources.
His Mag Sepharose Ni products are magnetic beads designed for efficient, small-scale purification/screening of histidine-tagged proteins from different sources. Histidine-tagged proteins are captured using immobilized nickel ions followed by collection of the beads using a magnetic device. You can use magnetic beads to simplify sample handling during small-scale purification.
His Mag Sepharose Ni delivers:
High-capacity small-scale purification of histidine-tagged proteins from different sources
High purity and yield
Easy parallel screening of histidine-tagged proteins with high repeatability
Scalability—Simple capture of histidine-tagged proteins from small or large sample volumes (low microliter to milliliter scale)
The beads are available in three pack sizes: 2 × 1 ml 5% medium slurry, 5 × 1 ml 5% medium slurry, and 10 × 1 ml 5% medium slurry—a 1 ml 5% medium slurry is the same as 50 μl sedimented medium. A 1 ml medium slurry is sufficient for 5 purification runs according to the recommended protocol.
Together with MagRack 6, a separation tool for handling the beads in microcentrifuge tubes, up to six samples can be processed in parallel. You can easily screen a larger number of samples in parallel with high throughput on a robotic device.
The magnetic bead format has excellent properties for small-scale experiments. The high density of the beads allows rapid capture by magnetic devices while the visibility of the beads ensures reliable collection of the bound histidine-tagged proteins in the purification procedure. The characteristics of the media are summarized in Table 1. All the products are provided with protocols optimized for purification of histidine-tagged proteins.
Tabel 1. Characteristics of His Mag Sepharose Ni
Matrix: Highly crosslinked spherical agarose (Sepharose) including magnetite
Metal icon capacity: 21µmol Ni2+/ml medium
Binding capacity: approx. 50mg histidine-tagged protein/ml medium (approx. 500µg/purification run)
Particle size: 37 to 100µm
Working temperature: Room temperature and + 4ºC
Storage solution: 20% ethanol, 5% medium slurry
Storage temperature: Room temperature
GE Healthcare Europe GmbH
Dr. Michael Walther EMEA Marketing Manager Electrophoresis
T + 49 2293 816 749 M + 49 173 574 9413 F + 49 5564 204 593
Efficient testing of the pro- or anti-angiogenic capacity of substances and drugs demands for standardized and easy-to-use test systems that deliver reproducible data quickly.
PromoCell provides four different ready-to-use angiogenesis cell model systems. Each of them allows the simulation of angiogenesis processes, by detailed in vitro recapitulation of the biological events involved in de novo vessel formation and remodelling: proteolytic activity, migration, proliferation and tube formation.
The PromoCell 3D Angiogenesis Kit examines all process together. It contains a collagen matrix with embedded endothelial cell spheroids and arrives ready-to-use in your lab. The only thing you need to do in order to start the assay is to add your test substance. Following proper stimulation de novo vessel formation occurs within two days, visible by sprouting of tubes into the surrounding matrix. The number and length of the sprouts corresponds to the pro- or anti-angiogenic potency of the test substance. Evaluation of the results is straightforward and easy, and staining procedures are not necessary.
To investigate the separate events of angiogenesis in a simulated in vitro environment, the PromoCell Transmigration Kit, the PromoCell Invasion Kit and the PromoCell Planar Migration Assay are useful platforms.
The PromoCell Transmigration Kit takes advantage of the ability of endothelial cells to migrate along cytokine gradients in a concentration-dependent manner, allowing for the measurement of the chemotactic potential of a test substance.
The PromoCell Invasion Kit is suitable for in vitro simulation of the proteolytic activity of endothelial cells as well as investigating directed migration. Hence, the impact of test substances on one or both of the aforementioned parameters can be easily monitored.
The PromoCell Planar Migration Assay is a 2D test platform for rapid evaluation of the influence of test substances on the migratory response of endothelial cells.
His Mag- and Ni Sepharose excel are immobilized metal ion affinity chromatography (IMAC) media (resins) designed for capture and purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants.
Nickel ions are very strongly bound to both media, enabling direct loading of large sample volumes without removing agents that normally would cause metal ion stripping. Both media significantly simplify and speed up the workflow and are excellent choices when working with purification of histidine-tagged proteins secreted from eukaryotic cells.
Increase target protein yield and decrease degradation through reduced and simplified sample handling
Choose between several different formats for screening and preparative purification of histidine-tagged proteins
IMAC purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants has traditionally been challenging because of incompatibility between the IMAC medium and the cell culture medium. The cell culture medium strips the immobilized metal ions from the IMAC medium during sample loading, resulting in low or no binding of the target protein. The purification has been further complicated by the fact that the target protein concentration is often low. This requires the use of large sample volumes, which in turn might lead to increased metal ion stripping. Considerable sample pretreatment has been required to overcome these problems, an example being buffer exchange by diafiltration in combination with concentration procedures. Such pretreatment is time consuming, potentially harmful to sensitive proteins, and can cause unnecessary loss of target protein.
Featuring proven bio-decontamination technology and flexible design, this new system offers significant efficiencies for cGMP pharmaceutical manufacturing.
By ensuring a controlled aseptic test environment, the Bioquell QUBE helps to minimise the risk of unnecessary, and costly, batch rejection due to false positive sterility test results.
The new workstation has been developed as a cost-effective, modular, barrier separation system. It incorporates Bioquell’s unique hydrogen peroxide vapour (HPV) technology to offer a rapid bio-decontamination cycle for fast and efficient transfers. HPV consistently delivers a high-level 6-log reduction in bioburden, validated by high quality HPV-optimised biological indicators. The Bioquell QUBE has been designed to accommodate a fully fitted or bench mounted sterility test unit, including the market leading Merck Millipore Steritest system. An EU GMP Grade A / ISO 5 compliant environment is ensured by both uni-directional airflow and unique isolator technology.
The intelligent modular design offers flexible configurations. If space is limited, a single module can be used. Conversely, if there is a need for high volume throughput, a three chamber workstation can be installed.
The Bioquell QUBE is the latest edition to the company’s portfolio of solutions for sterility assurance in cGMP pharmaceutical and biotechnology processes, backed by over 200 years of decontamination experience. Explore the new generation of hydrogen peroxide vapour (HPV) bio‑decontamination technologies from Bioquell at www.bioquell.com/bioquell-qube/
AMSBIO announces the introduction of CloneCatcher™ Gold DH5G Electrocompent E. coli cells - a new product that exhibits extremely high electroporation efficiencies.
The new product exhibits extremely high electroporation efficiencies that approach the theoretical maximum of 3.4 x 1011 cfu/µg pUC19 DNA. Transferring exogenous DNA into E. coli is a standard laboratory method for cloning genes and constructing cDNA, genomic and epitope libraries. The limiting factor in many library-screening efforts or multiple fragment ligations is the efficiency by which DNA can be introduced into E. coli. Electroporation is one method to efficiently introduce DNA into E. coli. CloneCatcher™ Gold cells meet the needs of scientists that are seeking to find rare clones or are building complex or metagenomic libraries. CloneCatcher™ Gold significantly outperforms all other commercially available Electrocompetent Cells when introducing ligated DNA. When the amount of input DNA is limited, CloneCatcher Gold enhances your likelihood of success. CloneCatcher™ Gold requires less topoisomerase-loaded vector and therefore can reduce your overall operating costs. Supplied in convenient single-use tubes - CloneCatcher™ Gold can also enhance your productivity as the whole protocol for use takes only 10 minutes.
For further information on CloneCatcher™ Gold please visit our Website.
rProtein A/Protein G GraviTrap includes ten ready-to-use columns prepacked with 1 ml of a mixture of 50% rProtein A Sepharose Fast Flow and 50% Protein G Sepharose 4 Fast Flow.
rProtein A/Protein G GraviTrap includes ten ready-to-use columns prepacked with 1 ml of a mixture of 50% rProtein A Sepharose Fast Flow and 50% Protein G Sepharose 4 Fast Flow.
•Prepacked gravity-flow columns provide convenience, save time, and increase the reliability of your results.
•Purification of antibodies in 30 min allows you to quickly obtain and store your target protein in order to avoid the inherent dangers of protein instability and degradation associated with slow and long purification runs.
•High purity and recovery with purity > 95% in one step and recovery typically 70% to 80%.
•High binding capacity such as Protein A/Protein G GraviTrap ~ 35 mg human IgG/column.
rProtein A/Protein G GraviTrap are prepacked gravity-flow columns designed for fast and efficient manual purification of monoclonal and polyclonal antibodies, antibody fragments from cell culture supernatant, and biological fluids. The antibodies are simply captured with high specificity on protein A and protein G ligands in gravity-flow columns. You do not need any other instrument with this protocol because the entire process relies on the flow of gravity. The yield varies from 20 to 50 mg of purified antibodies depending on the ligand you use. The columns are reusable up to five times.
PromoKine offers a huge and continuously growing range of recombinant cytokines, chemokines, growth factors, soluble receptors and CD antigens (human, mouse, rat).
PromoKine offers a huge and continuously growing range of recombinant cytokines, chemokines, growth factors, soluble receptors and CD antigens (human, mouse, rat) that are produced in E. coli, yeast, insect, plant or mammalian cells. They are highly purified, biologically active and stable and are available in standard and very economic bulk sizes.
PromoKine has also extended its range of animal-free cytokines and growth factors that are produced in plants and are thus free of animal components, serum, endotoxins and antibiotics. These are also free of human, animal or bacterial infectious agents as well as other endogenous mammalian agents that might cause unwanted cell responses. They show very low proteolytic, pyrogenic and inflammatory activity compared to other commercially available proteins. Moreover, they retain proper eukaryotic folding and glycosylation, resulting in increased stability and biological activity.
In addition, PromoKine offers a huge variety of high-quality antibodies and ELISAs for the analysis of cytokines and growth factors as well as other interesting targets in cell biology.
TiO₂ Mag Sepharose is magnetic beads designed for enrichment of phosphopeptides from tryptic digested protein samples
Magnetic beads for selective capture and enrichment of phosphopeptides
Easy to use - visible and dense magnetic beads
Simple protocol with elution conditions optimized for MS analysis
Parallel handling of samples - six samples can be processed in less than one hour
TiO2 Mag Sepharose magnetic beads are based on titanium dioxide (TiO2) chromatography and are designed to simplify capture and enrichment of phosphopeptides. TiO2 has high affinity for phosphopeptides and is an efficient method for enrichment of phosphopeptides from complex samples
TiO2 Mag Sepharose improves ease-of-use in sample preparation of phosphopeptides. The magnetic bead format has excellent properties for small-scale experiments. The high density of the beads allows rapid capture by magnetic devices while the visibility of the beads ensures reliable collection of all bound target peptides
Together with MagRack 6, a separation tool for handling the beads in microcentrifuge tubes, up to six samples can be processed in parallel