Amaxa cell line optimization nucleofector kit lonza bioscience. The nucleofector technology, comprising nucleofection process, nucleofector device, nucleofector solutions, nucleofector. The nucleofector ii device is the basic device for single cuvette applications. Jun 28, 2007 nucleofector device amaxa, aad1001 rat neuron nucleofector kit amaxa, vpg1003 or mouse neuron nucleofector kit amaxa, vpg1001 containing cuvettes for the nucleofection reaction, plastic. The first is the nucleofector tm device, which offers a unique portfolio of electrical parameters, displayed as distinct programs that are adapted to the requirements of specific cell types. The ratio of nucleofector solution to supplement is 4. For preparing aliquots, mix nucleofector solution and supplement in a ratio of 4. Transfection efficiency of freshly isolated human b cells 24 hours post nucleofection.
Use of amaxas nucleofector technology requires a license from amaxa gmbh. Download scientific diagram efficiency of cellular transfection between amaxa. Overview note bacteria programs can be only selected via the cell type list mode. Nucleofector technology offers high flexibility within applications. Competitive dna transfection formulation via electroporation. It employs a nonviral method based on an optimized combination of electrical parameters and celltype specific solutions. Jun 10, 2011 the amaxa nucleofector amaxa gmbh, cologne, germany is an electroporation device combined with a set of special transfection buffers that greatly improves the transfection efficiency and. The amaxa nucleofector uses a novel nonviral transfection technology to introduce nucleic acids both dna and rna into stem cells, primary cells, and cell lines that are difficult to transfect. Transfecting human neural stem cells with the amaxa.
Optimized protocols that are provided with the nucleofector kits which. Genomic editing, nucleofection, and the creation of next. Manufacturer specifications amaxa 4dnucleofector system, lonza. The condition that results in the highest transfection efficiencies with the lowest amount of cell death is chosen for the rest of the studies. Place the cuvette into the nucleofector, rotate the wheel clockwise, and select program. Customers include pharmaceutical and biotechnology companies, academic and government institutions.
Nucleofector technology accelerating cancer research. With an extremely fast plate processing time of one minute, it is. Each solution and supplement is individually developed for every primary cell type. Amaxa 4dnucleofector protocol for stimulated human t cells required material note please make sure that the entire supplement is added to the nucleofector solution. Optimizing these methods is possible, but many variables have to be taken into account. It is suggested that you try all 5 programs a23, a27, a, a12, b16 with each nucleofection solution initially to determine the optimal program. With lonzas researchuse human bone marrow and peripheral blood immune cells you dont have to hassle with finding donors and isolating specific cell.
Due to this independency from cell proliferation, it allows for efficient transfection of even nondividing primary cells like resting t cells or neurons and also speeds up expression. Oct202015 just being cheap and trying to save money for the lab since we use the genepulser cuvettes for li electroporation already. Amaxa 4dnucleofector protocol for human stem cell line h9. The comprehensive way in which nucleofector programs and cell typespecific solutions are developed enables nucleic acid substrates delivery not only to the cytoplasm, but also. Amaxa 4dnucleofector protocol for shsy5y 2 required material note please make sure that the entire supplement is added to the nucleofector solution. Perform the nucleofection as per manufacturers instruction using an amaxa nucleofector ii device. Please try all 5 nucleofector programs initially to determine the most appropriate one for your neuron cell type for all subsequent experiments. Electroporation, nucleofection and plasmids researchgate, the professional. I want to transfect murine hematopoietic progenitors with a cas9encoding. It is used to set up, control and save the results of an experiment. It enables highly efficient, transfection of primary cells, stem cells, neurons, and cell lines that have traditionally been difficult to transfect via electroporation and other nonviral. It is important pinpointing that the transfection of nonreplicative forms trypomastigote is less efficient than that of the epimastigotes. The first set of experiments is comprised of 18 reactions. Does anyone have experience with amaxa nucleofector based.
The controlling unit of the 4d nucleofector tm system is suited to handle various fuctional units for efficient transfection of hard to transfect cell lines and primary cells in different formats. Ingenio cuvettes for the ezporator electroporation system and lonzaamaxa nucleofector ii2b devices ingenio cuvettes for the ezporator electroporation system and all conventional electroporators such as biorad and harvardbtx ingenio cell droppers. This is an extract of the amaxa biosystems protocol vs. Select nucleofector program insert cuvette press start button x expression detected as early as 38 hours 6 our product family a technical comparison device nucleofector 2b 96well shuttle 4d nucleofector ht nucleofector. Initially please try all 5 nucleofector programs with both nucleofector solutions to determine the most appropriate nucleofection condition for your specific human stem cell type. This manual describes the ht nucleofector software and is valid for ht nucleofector software. Amaxa nucleofector protocol for rat oligodendrocytes. Optimized protocol for efficient transfection of dendritic. Amaxa 4dnucleofector protocol for human stem cell line h9 required material note please make sure that the entire supplement is added to the nucleofector solution. Below, you will find the composition of the buffers and the supplement solutions.
Highefficiency transfection of mammalian neurons via. Oct 26, 2010 easy and effective procedure of nucleofector technology by lonza, usa for more information, please visit. Lonza bioscience is a leader in several highvalue segments of the lifescience industry, providing products and services to support the discovery and commercialisation of human therapeutics. Amaxa general nucleofection protocol for transformation of bacteria application this protocol enables you to optimize the nucleofection conditions for the transformation of different bacteria strains. Nucleofection condition with the highest efficiency and lowest mortality is selected. The results of this first step are displayed in figure 1a. Purchasers are granted a nonexclusive, nontransferable license for the use of amaxas nucleofector. The electrical parameters are different from any other.
Amaxa nucleofector rensselaer center for biotechnology. Does anyone have experience with amaxa nucleofector based transfection of murine hematopoietic progenitor cells. Nucleofector supplement is added to the nucleofector solution it is stable for three months at 4c. Use ingenio with amaxa nucleofector device transfection. Transfer the cells to 1 ml of hspc culture medium without penicillinstreptomycin in 48well plate see note 5. Use of amaxa s nucleofector technology andor related software requires a license from amaxa ag. Biomolecular analysis involves the characterization of polymeric biomolecules such as dna, rna, protein, carbohydrates and lipids as well as single biomolecules. Nucleofector ii general description a single cuvette based system used for the transfection of cell lines and primary cell that are hard to infect with various substrates including dna vectors or sirna olignucleotides. Ingenio electroporation products for high efficiency. Transfer the cells to a 37 c incubator and culture for 24 h. It allows efficient transfection of hardtotransfect cell lines and primary cells for transfection in lowthroughput format with. Purchasers are granted a nonexclusive, nontransferable. The nucleofector device is an apparatus which delivers unique electrical parameters. The controlling unit of the 4d nucleofector tm system is suited to handle various fuctional units for efficient transfection of hardtotransfect cell lines and primary cells in different formats.
Please note that the amaxa nucleofector technology is not intended to be used for diagnostic purposes or for testing or treatment in humans. To determine the nucleofection program that would yield the highest effectiveness, we used the following seven programs, as outlined in the amaxa nucleofector optimization protocol. Optimization guidelines the initial optimization experiment is comprised of 6 reactions. Enhanced nucleic acid delivery using conventional electroporation devices that provide. Amaxa 4dnucleofector optimization protocol for cell lines. Use of amaxas nucleofector technology andor related software requires a license from amaxa gmbh. Transfecting human neural stem cells with the amaxa nucleofector. Ppt 4dnucleofector powerpoint presentation free to. Having tested various fibroblastoid cell types, high transfection efficiencies could be achieved using one of the programs indicated below. The ingenio solution will yield results similar to those obtained using the.
Nucleofection conditions can be performed with the help of our scientific. Nucleofection is an electroporationbased transfection method which enables transfer of nucleic acids such as dna and rna into cells by applying a specific voltage and reagents. Detection of pathogenic antigen by naive dcs is through pattern recognition receptors prrs which are able to recognize specific conserved structures referred to as pathogenassociated molecular patterns pamps. The comprehensive way in which nucleofector programs and cell typespecific solutions are developed enables nucleic acid substrates delivery not only to the cytoplasm, but also through the nuclear membrane and into the nucleus, transfection into the nucleus, hence nucleofector. Effective gene delivery to trypanosoma cruzi epimastigotes.
The nucleofector ii2b transfection device is the single cuvette based system that has been used in research labs since 2001, and formerly known as the amaxa device. Are biorad genepulser cuvettes compatible with a lonza nucleofector 2b system. Genomic editing, nucleofection, and the creation of nextgeneration cellbased assays. None of the equipment posted for sale on is owned by medwow, should you have any questions regarding a specific item, please direct them to the appropriate seller by making use of the available communication channels on the items page. Aug 19, 2019 transfection technology expert amaxa will lead the consortium in its quest to develop uht devices that can effectively deliver genetic material such as dna or sirna small interfering rna into primary cells using amaxa s nucleofector technology before allowing subsequent cell screening in third party equipment. Aug 29, 20 transfection of lncap cells with dna plasmids was performed using amaxa nucleofector kit r from lonza. Cryopreserved ampule of human umbilical vein endothelial cells huvec without vegf. The optimization process involves subjecting cells to various combinations of electrical parameters termed programs in the nucleofector device and different nucleofector solutions.
Transfection using the nucleofector is a patented commercial technique requiring special buffers and programs, the constituents of which are a secret. General protocol for nucleofection of adherent cell lines. Nevertheless, we developed in house methods for ecm 630 electroporator machine. Amaxa basic nucleofector kit for primary mammalian. Transfer the cells to 1 ml of hspc culture medium without. Electroporation by nucleofector is the best nonviral. Trouble with amaxa nucleofection in my lab we need to deliver dna to b cells. Two different cell line nucleofector solutions l and v are tested in combination with 7 different nucleofector programs plus 2 controls. With the amaxa nucleofector, use their recommended program or a preferred userestablished protocol. Amaxa 4dnucleofector protocol for stimulated human t. The components of the amaxa nucleofector technology the nucleofector kits nucleofector kits contain a specifi c nucleofector solution and supplement, specifi ed cuvettes, pipettes, and the pmaxgfp control vector. Jul 08, 2011 dendritic cells dcs can be considered sentinels of the immune system which play a critical role in its initiation and response to infection 1. A012, a0, a023, a027 and b016 a12, a, a23, a27 and b16 for nucleofector i device.
The nucleofector tm technology uses a specific combination of optimized electrical parameters and cell typespecific solutions which enables transfer of a molecule directly into the cells nucleus. The ingenio electroporation kits and solution can be used with any conventional electroporation instrument, including the amaxa nucleofector ii system. Download document genome editing using nucleofector tm technology technical reference guide providing a brief introduction to genome editing tools and technical tips for zfn, talen or crispr delivery using the nucleofector. Manufacturer specifications amaxa 4d nucleofector system, lonza please note. They are resistant to easy transfection methods and lately we have been having issues with our outdated retroviral system. Nucleofection, also referred to as nucleofector technology, was invented by the biotechnology company amaxa. Both dna and rna are efficiently transfected into both the. Are biorad genepulser cuvettes compatible with a lonza. Amaxa nucleofector protocol for rat oligodendrocytes for primary rat oligodendrocyte precursor cells isolated from forebrains of newborn rats p1 and cultivated for 17 days before nucleofection. Ht nucleofector software manual 1 introduction the ht nucleofector software is the graphic user interface required to operate the ht nucleofector system. Handmade solutions for amaxa lonza nucleofector facebook. Amaxa 4dnucleofector protocol for mouse embryonic fibroblasts mef required material note please make sure that the entire supplement is added to the nucleofector solution. Efficiency of cellular transfection between amaxa nucleofector. It is also fast, so for easy to use, preconfigured kits that work especially well in primary samples, including patient sample material, it is highly recommended.
Biomolecular analysis core biomolecules form the basis of life. Our solution is an improved electroporation technology, the nucleofector tm technology, originally introduced into the market by legacy amaxa in 2001. May 04, 2004 overall, the amaxa nucleofector system and kits are extremely convenient, easy to use, and beneficial for primary cell research where nonviral gene transfer is required. The comprehensive way in which nucleofector programs and cell type. Purchasers are granted a nonexclusive, nontransferable license for a limited use of. Amaxa 4dnucleofector protocol for human umbilical vein cells huvec required material note please make sure that the entire supplement is added to the nucleofector solution. Ingenio electroporation solution competitive comparison. The amaxa nucleofector uses a novel nonviral transfection technology to introduce nucleic acids both dna and rna into stem cells, primary cells, and cell lines that are difficult to. A free powerpoint ppt presentation displayed as a flash slide show on id. Competitive electroporation formulation for cell therapy. Nucleofection is a technology based on the momentary creation of small pores in cell membranes by applying an electrical pulse. Optimized protocol for cell line optimization nucleofector kit 2 optimization guidelines step 1.
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