Neurobiology Research Tools for Biologically Relevant Results

Transcription

1 BioResearch Neurobiology Research Tools for Biologically Relevant Results Neurobiology Research Tools

2 BioResearch Neurobiology Research Tools Avoid Research Roadblocks and Take a Direct Path to Results " Sourcing cells from specific species and brain regions requires a lot of my time and effort." SOLUTION: Select from Lonza s range of well-characterized and ready-to-use primary cells. Simply thaw and culture. Page 5 Primary Cells and Media "Non-viral transfection doesn t give me the transfer efficiency I m looking for." SOLUTION: Our non-viral Nucleofector Technology delivers 70 % gene transfer efficiency and high expression levels. Page 9 Transfection 2

3 " I can t find reliable, ready-to-use assay tools for protein expression, cell proliferation, or cytotoxicity." SOLUTION: Choose from our variety of neurology-specific assay tools, including ready-to-use protein gels, and bioassay kits to measure cell proliferation and cytotoxicity. Page 14 Cell Analysis " I need to develop co-culture systems in 3D with animal neural cells" SOLUTION: With Lonza s broad animal neural cells portfolio combined with RAFT 3D culture system, you can develop co-culture systems that more closely represent an in vivo model. Page 13 Drug Discovery 3

4 BioResearch Neurobiology Research Tools Your Complete Neurobiology Tool Kit Streamline your neurobiology workflow by choosing from convenient, innovative research tools that have been designed and tested to work together. Lonza solutions provide biologically relevant results, from high-quality primary cells, through efficient transfection technology, to expert analysis tools. Primary Cells and Media Transfection Drug Discovery Cell Analysis Ready-to-use cryopreserved Clonetics or Poietics Primary Neural Cells: Astrocytes (mouse, rat, or human) Cerebellar granule cells (rat) Cortical neurons (mouse or rat) Dorsal root ganglia (rat; embryonic or neonatal) Hippocampal neurons (rat or mouse) Hypothalamus neurons (rat) Neural progenitor cells (human) Retinal neurons (rat) Striatum neurons (mouse or rat) Optimized Clonetics Primary Neural Cell Growth Media: PNGM or PNGM -A BulletKit Media for embryonal or adult primary animal neurons AGM BulletKit for human and animal astrocytes NPMM or NPDM BulletKit Media for neural progenitor cell growth or differentiation Nucleofector Technology with optimized protocols for primary cells, including: Hippocampal neurons Cortical neurons Dorsal root ganglia Cerebellar granule cells Astrocytes Oligodendrocytes Neural stem cells Also optimized for numerous cell lines, including: PC-12 SH-SY5Y Neuro2A RAFT 3D Culture System for advance cell culture solutions Blood brain barrier models Cells on Demand Services: Custom transfection services Custom cell isolation services Cell Proliferation and Cytotoxicity BioAssay Kits: ViaLight Kit for measuring cell proliferation and cytotoxicity ToxiLight Kit for nondestructive measurement of cytotoxicity Protein Expression Analysis: PAGEr Precast Protein Gels for Western blotting 4

5 Primary Neural Cells and Growth Media Access neural cells when you need them. Choose from Lonza s extensive range of isolated primary neural cells and culture them in media that we have already optimized and tested to give you the best results. You will never have to put research on hold for animal pregnancies again. Simply store Lonza cells and thaw them as needed. Cryopreserved Clonetics Primary Neurons " Sourcing cells from specific species and brain regions requires a lot of my time and effort." SOLUTION: Select from Lonza s range of well-characterized and ready-to-use primary cells. Simply thaw and culture. Choose from our broad selection of high-quality, high-yield cultures of dissociated primary animal neurons that are: Tested for neuron-specific markers Guaranteed to perform in culture Guaranteed free of mycoplasma and bacteria Tested to ensure they will perform after shipping Lots can be reserved to repeat experiments using cells from the same batch Rat Mouse (CD1 or C57) Hippocampal neurons PNGM Cortical neurons PNGM Striatum neurons PNGM Recommended Media Dorsal root ganglia (embryonic or neonatal) PNGM Cerebellar granule cells PNGM -A Hypothalamus neurons PNGM Retinal neurons PNGM Microglia DMEM 4.5 g/l Glucose w/ L-Gln, 500 ml Neural networks of cultures (7 DIV) prepared from (A) fresh and (B) cryopreserved rat hippocampal neuronal cells immunostained with the specific neuron marker anti-pgp9.5. Applications Transfection Evaluating electrophysiological properties, neurotransmitters, and receptor function Drug screening Research into ion channels, intracellular transport, and neurotoxicity 5

6 BioResearch Neurobiology Research Tools Clonetics PNGM Primary Neuron Growth Media Cryopreserved Clonetics Primary Astrocytes Clonetics PNGM gives you a proven alternative to Neurobasal and B27 for long-term culture of embryonic or adult rat and mouse neurons: Serum-free media in BulletKit Format Guaranteed to perform with all Clonetics Primary Animal Neurons Batch tested with primary neurons and a pre-screened B27 equivalent supplement Applications Culture embryonic primary neurons in PNGM BulletKit Media. Culture adult primary neurons and cerebellar granule cells in PNGM -A BulletKit Media Growth of Rat Cortical Neurons in Different Media Relative RLU (normalized to control) No Cells Neurobasal PNGM Clonetics Rat Cortical Neurons were seeded at 40 % recommended density and cultured in PNGM BulletKit Media and a competitor media. After 6 DIV, viability was determined by ViaLight Plus Assay. Values were normalized to no cell control. Improve your workflow with cryopreserved, high-quality human, and animal glial cells. There is no need for animal care or glial cell isolation with these products, which are: Passaged once after isolation Batch tested for growth characteristics and morphology (GFAP) Guaranteed free of mycoplasma Able to be reserved by lot so you can repeat experiments using cells from the same batch Rat Mouse (CD1 or C57) Human Astrocytes Hippocampal astrocytes Cortical astrocytes Striatum astrocytes Cx-Hi-Cp mixed Astrocytes Cx-Hi-Striatum mixed Astrocytes Applications Transfection Pharmacology Drug screening Research into neurogenesis, cell physiology, Alzheimer s disease, Parkinson s disease, spinal cord injury, astrocyte-mediated neurotoxicity, ion channels, electrophysiology, and patch clamping Clonetics AGM Primary Astrocyte Growth Media Culture human and animal astrocytes with serum-free media, in BulletKit Format. This media has been tested and guaranteed to perform with all Clonetics Primary Astrocytes. 6

7 Cryopreserved Poietics Neural Progenitor Cells Safeguard your experiment with progenitor cells that are guaranteed to differentiate into a mixed population of neuronal cells that express stipulated markers on laminin-coated plates. Cryopreserved in primary passage as spheroids, our high-quality cells are tested: Positive for ß-tubulin III (neurons) and GFAP (astrocytes) Negative for HIV-1, Hepatitis B and C, mycoplasma, bacteria, yeast, and fungi Together with media and reagents to ensure they perform optimally as a system Sold under license from StemCells, Inc. US patents 5,968,829 and 5,851, 832. Applications Transfection Drug development Research into neurotoxicity, neurogenesis, electrophysiology, CNS function, and neurotransmitter disorders Normal human neural progenitor cells stained for ß-tubulin III and GFAP. Poietics Neural Progenitor Media Choose from two serum-free media that are specially formulated to support growth, expansion and differentiation of normal human neural progenitor (NHNP) cells: For optimal neural progenitor cell growth and differentiation, there s the NPMM Neural Progenitor Maintenance Medium BulletKit For optimal neural progenitor differentiation, we have NPDM Neural Progenitor Differentiation Medium BulletKit Indirect immunofluorescence staining of Clonetics Normal Human Astrocytes for glial fibrillary acidic protein (GFAP). 7

8 BioResearch Neurobiology Research Tools Primary Cell Case Study: Co-cultures for Neuropathology Research Co-culture of cryopreserved rat primary cortical and striatal neurons. B. Tinner-Staines, et al Society for Neuroscience; Abstract No Background Most primary neuronal cell cultures use neurons from a single brain region but this does not provide an adequate model to study neuropathologies such as Alzheimer s or Parkinson s diseases. De-afferented neurons demonstrate biochemical and physiological abnormalities that limit scientific study and preclude drug screening. To model the circuitry associated with different neuropathologies, relevant neuronal cell types must be co-cultured in vivo. It is difficult to time and stage co-culture of freshly isolated neurons according to the normal developmental staging of cell connectivity. However, this study shows that cortical and striatal cryopreserved neurons in co-culture could provide viable models for examining questions of basal ganglia function and toxicology. Plating a combination of cryopreserved rat striatal and cortical neurons yields cultures characteristic of both cell types cultured individually, as shown by staining of cell soma (PGP 9.5) and neurofilaments (NF-160). Thawed cortical and striatal rat neurons were co-cultured for 21 days and stained with antibodies against vesicular GABA transporter (vgat; green) and dopamine receptor protein (DARPP-32, red). The presence of GABAergic innervation of DARPPpositive neurons gives evidence of crossinnervation between neurons. Method Conclusion Cryopreserved primary Clonetics Rat Cortical and Striatal Neurons were thawed, plated in tandem, and incubated for 735 days in vitro. Cultures were characterized using synaptic marker antibodies, receptor and second messenger proteins, and structural marker proteins. Results Results demonstrate that in vitro modeling of brain circuits can be easily accomplished by using a co-culture of cryopreserved primary neural cells. These facilitate co-cultures according to the normal developmental staging of cell connectivity because researchers do not need to coordinate isolations from different brain regions or embryonic developmental stages. Cortical and striatal co-cultures displayed characteristics of each individual neuron type. Furthermore, co-cultures showed nerve terminals expressing vesicular GABA transporter or glutamate transporter (latter not shown here), which is evidence of neuronal crosstalk, thus reflecting the in vivo situation. 8

9 Transfection of Primary Neural Cells or Cell Lines Nucleofection was the first efficient non-viral transfection technology for primary neurons. Its unique combination of electrical parameters with cell type-specific solutions and protocols gives you up to 70 % gene transfer and excellent cell viability. Now this innovative technology also allows you to transfect primary neurons or glial cells in adherence at later developmental stages. Nucleofector Technology " Non-viral transfection doesn t give me the transfer efficiency I m looking for." SOLUTION: Our non-viral Nucleofector Technology delivers 70 % gene transfer efficiency and high expression levels. Nucleofection gives you: High transfection efficiencies and cell viability Preservation of cell functionality A variety of device platforms for different cell numbers and throughput Nucleofection of primary neurons or glial cells in adherence, even after several days of culture Optimized protocols to save time and effort Choose the Nucleofection Platform that Suits Your Research Needs Advanced Platform 96-well Add-on High-throughput Platform Basic Device Device 4D-Nucleofector System 96-well Shuttle System 384-well Nucleofector System Nucleofector 2b Device Unit Throughput (samples per run) Low to medium (1 16) Low to high (1 96) High (384) Low (1) Reaction volume 20 µl µl 20 µl 20 µl 100 µl Electrode material Conductive polymer Conductive polymer Conductive polymer Aluminum Low cell numbers (20 µl) to to to High cell numbers (100 µl) DNA Vector amount/sample sirna amount/sample (concentration 2 nm 2 µm) μg (20 µl) 1 5 μg (100 µl) μg μg 1 5 μg pmol (20 µl) pmol (100 µl) pmol pmol pmol Adherent Nucleofection Compatibility with 96-well Shuttle System To find transfection data for your cell type of interest. 9

10 BioResearch Neurobiology Research Tools Transfection Efficiency and Cell Viability with Nucleofector Technology Efficiency* Viability* Kit for 4D-Nucleofector and 96-well Shuttle Systems (name of specific protocol) Kit for Nucleofector II/2b Device Primary Neural Cells Astrocytes, mouse 5565 % 6070 % P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Astrocytes, rat 6070 % 7080 % P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Dorsal root ganglia (DRG), chicken 2535 % P3 Primary Cell (Neuron, Basic) Chicken Neuron Dorsal root ganglia (DRG), rat 3545 % P3 Primary Cell (Neuron, Basic) Rat Neuron Neuron, cortical, rat 3070 % 4560 % P3 Primary Cell (Neuron, rat) Rat Neuron Neuron, hippocampal, chicken 4050 % P3 Primary Cell (Neuron, Basic) Chicken Neuron Neuron, hippocampal, mouse 5060 % P3 Primary Cell (Neuron, Basic) Mouse Neuron Neuron, hippocampal, rat 3070 % 4560 % P3 Primary Cell (Neuron, rat) Rat Neuron Neuron, hippocampal, rat adherent 3050 % 5080 % AD 1 Primary Cell (Neuron, Basic, AD) Not Applicable Neuron, cortical, rat adherent 3060 % 7090 % AD 1 Primary Cell (Neuron, Basic, AD) Not Applicable Neuron, cortical, mouse adherent 2070 % 7090 % AD 1 Primary Cell (Neuron, Basic, AD) Not Applicable Oligodendrocyte, rat 4050 % 5565 % P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Stem Cells Neural stem cell (NSC), mouse 7585 % Primary Cell Optimization Mouse NSC Neural stem cell (NSC), rat 4050 % Primary Cell Optimization Rat NSC Neuronal Cell Lines** AGN2a 6095 % Cell Line Optimization Cell Line R BV % 7095 % Cell Line Optimization Cell Line T C % 7080 % Cell Line SF Cell Line V H % 8090 % Cell Line SG Cell Line V IMR % 5565 % Cell Line SF Cell Line L Neuro2A 4785 % 8095 % Cell Line SF Cell Line V NG % 6095% Cell Line SF Cell Line V PC % 7585 % Cell Line SF Cell Line V SH-SY5Y 6085 % 4080 % Cell Line SF Cell Line V SK-N-AS 9096 % 82 93% Cell Line SF Cell Line V SK-N-MC 6095 % 3060 % Cell Line SF Cell Line V SK-N-SH 9599 % 5067 % Cell Line SF Cell Line V U-87MG 3545 % 8595 % Cell Line SE Cell Line T Primary cells marked blue have Lonza-validated optimized protocols. * Approximate ranges extrapolated from larger result collections, including Lonza and customer data ** This is only a selection of cell lines. For further neuronal cell lines and protocol guidance. 10

11 Adherent Nucleofection of Primary Neurons at Later Developmental Stages Transfect primary neural cells at any stage of culture, including late development, without putting them in suspension. Forget about the traditional limitations of electroporation-based methods with our new 4D-Nucleofector Y Unit that enables adherent Nucleofection: Pre- and post Nucleofection culture in 24-well culture plates Nucleofection of neuronal cells at any time point during this culture period, i.e. at a later developmental stage Transfection efficiencies up to 70 % combined with high viabilities Compatible with Clonetics Primary Animal Neurons Efficient adherent Nucleofection of neurons in 24-well culture plates. Mouse cortical neurons were seeded into poly-d-lysine coated 24-well plates (1 105 cells/well). After 6 DIV, cells were transfected with maxgfp Vector using the AD1 4D-Nucleofector Y Kit. One day post Nucleofection, cells were stained by MAP2 antibody (red) and analyzed by fluorescence microscopy for maxgfp Protein Expression. Cryopreserved dissociated rat DRG cells were thawed and cultured in 24-well plates for Nucleofection using 4D-Nucleofector Y Unit. DRG cell culture was transfected at 2 DIV and fixed 24 hours post Nucleofection (program EH-166). Neuronal networks are stained using anti Tuj-1 antibody (red; personal gift W. Staines). Transfected neurons and Schwann cells can be seen in green (maxgfp Protein). 11

12 BioResearch Neurobiology Research Tools Transfection Case Study: Parkinson's Research Are neural progenitor cells (NPCs) better suited for transplantation when transfected with neurotrophic factors? Cesnulevicius, et al Stem Cells; 24(12): Background Transplanting neuronal progenitor cells (NPCs) or dopaminergic neurons might have therapeutic potential for neurodegenerative diseases, such as Parkinson s. However, transplanted cells show limited survival and are hard to identify in situ. This study investigated whether NPCs transfected with neurotrophic factors survived and matured more successfully after transplantation. The challenge was to find a non-viral transfection method that provides high efficiency without morphological or functional changes. Conclusion Cells transfected by Nucleofection displayed an unaltered morphology, could be differentiated into neurons, and were viable after transplantation. By delivering higher expression levels, Nucleofection could help to further investigate the therapeutic potential of NPCs. % Transfection Efficiency Electroporation Lipofection Nucleofection DsRed EGFP Method To identify the best method for transfecting NPCs derived from ventral mesencephali of rat brain, the study tested electroporation, lipofection, and Nucleofection. Ventral mesencephalic progenitor cells from rat brain were transfected with two different plasmids expressing DsRed or egfp using conventional electroporation, lipofection, or Nucleofection. Results Nucleofection was the most efficient method for NPCs, achieving a transfection rate of more than 45 %. 12

13 Drug Discovery RAFT 3D Cell Culture System 3D cell culture differs significantly from the traditional 2D culture that most researchers have been using since the past decades. There is a trending shift both in academia and industry to personalized research solutions and more in vivo like models to understand cell behavior. This is fueling the growing market need for better solutions in 3D cell culture such as RAFT 3D Cell Culture System. An important differentiation of the RAFT System to other 3D platforms or to 2D culture methods is its ability to engineer tissue-like 3D cultures. This is especially needed for certain applications such as development of in vitro liver fibrosis models or corneal models. With RAFT System, cells can be cultured within a high-density collagen scaffold, or on top, or both. The addition of permeable membrane cell culture inserts provides other extensions to the system allowing the generation of barrier models including air-lift models. I need to develop co-culture systems in 3D with animal neural cells SOLUTION: With Lonza s broad animal neural cells portfolio combined with RAFT 3D culture system, you can develop co-culture systems that more closely represent an in vivo model. RAFT Cultures can be created in less than an hour using simple protocols and standard labware! RAFT Kits are available in a choice of 24-well, insert-well and 96-well formats. RAFT System has already been used to successfully generate 3D cultures in a number of research areas including oncology, toxicology, barrier modeling, dermal research and pulmonary research. Contact your local sales representative to learn how RAFT System can support your experiments. RAFT 3D Culture System consists of RAFT Reagent Kit and Absorbers for 96-well, 24-well and trans-well inserts. Cells on Demand Services The advancement of assay technologies has enabled the use of live cells in over 50% of high throughput screening. To meet this growing need, Lonza research solutions has leveraged its decades of cell culture and transfection experience to provide high quality cell products and services. Cells on Demand Transfection Services Access Lonza s transfection expertise to achieve high transfection efficiencies and viabilities in almost every cell. Benefits include: Receive validated stable clones quickly Stress-free optimization of Nucleofection Protocols for your cell type of interest Cells on Demand Cell Culture Services Avoid delays in obtaining vital cell cultures. Just select the cell types and format and our cell culture experts will do the rest. Easily move to primary cells with our primary cell expansion service Avoid the aggravation of tissue acquisition, failed isolations, and low yields with our primary cell isolation service We can establish RAFT 3D Cell Culture Systems tailored to your needs within 96-well or 24-well formats 13

14 BioResearch Neurobiology Research Tools Cell Analysis Free your gene expression experiments from biases and interference with integrated research tools for every step of your workflow. Lonza solutions provide biologically relevant results from cell proliferation and cytotoxicity assays, to sample preparation and qpcr, all the way through protein confirmation. " I can t find reliable, ready-to-use assay tools for mrna, protein expression, cell proliferation, or cytotoxicity." Precast PAGEr Protein Gels for Consistent, Reliable Western Blotting PAGEr Gels are easy to use precast protein mini-gels that offer sharp resolution and consistent protein transfer. Each lot is functionally tested and shipped fresh every time, for guaranteed shelf life and performance. PAGEr Gels give you: Sharp resolution for crisp separation of proteins kda Marked sample lanes and simple twist open design Compatibility with most chambers Multiple well formats and gel concentrations Tris-Glycine buffer for traditional Laemmli separation Applications SDS-PAGE Native PAGE Western blotting Antibody screening SOLUTION: Choose from our variety of neurology-specific assay tools, including qpcr arrays, ready-to-use protein gels, and bioassay kits to measure cell proliferationand cytotoxicity. Comb Options and Well Volumes 2D well 550 μl sample, or 7 cm IPG strip, 12 μl marker 10 well 32 μl 8+1 well* 30 μl sample 12 μl marker 16 well 14 μl *Compatible with multichannel pipettes 12 well 20 μl 17 well* 14 μl 14

15 ViaLight Plus Kits Measure cell proliferation and cytotoxicity by using stable bioluminescence to detect adenosine triphosphate (ATP): Process and analyze a 96-well plate in less than 15 minutes Detect as few as ten cells; allowing for lower seeding densities and more assays Easily perform scalable automation with our simple, no-shake protocol Add two reagents directly to your culture and read luminescence Expand your dynamic range to five decades, in adherent or suspension cultures Run this test on a variety of luminometers or scintillation counters Avoid radioactive or toxic materials ToxiLight Kits Check a compound s cytotoxic effects non-destructively by measuring adenylate kinase (AK) leakage from damaged cells: Generate results from as few as 10 cells Eliminate the need to lyse by monitoring cytotoxicity from supernatant Simply add a single reagent directly to cells, or to a supernatant aliquot Process and analyze a 96-well plate in less than 10 minutes Freeze supernatants and return to your work later without losing AK activity /vialight EC 50 Data Generated Using ViaLight Plus Shows Consistency Over Time Identity Dose-dependent Activities in Cells 0h EC μg/ml 4h EC μg/ml 1h EC μg/ml 3h EC μg/ml 2h EC μg/ml 5h EC μg/ml ml HepG2 cells were incubated with the alkylating agent Mitomycin C for 48 hours and the assayed using ViaLight Plus. The experimental values are the mean of eight replicant samples read every hour over a 5 hour period. The EC values remain consistent over the 5 hour read period. Comparison of ViaLight Plus and ToxiLight Kits using HUVECs dosed with camptothecin. The ATP levels indicated by the ViaLight Plus RLUs reduce steadily in a dose-dependent manner. At the lower drug doses, the AK released from the cells is relatively low compared with that of the control, only increasing dramatically at the highest drug doses. 15

16 BioResearch Neurobiology Research Tools Ordering Information Cat. No. Description Size RAFT 3D Cell Culture System 016-1R10 RAFT 96-well Bundle Kit, contains (016-0R94 & 016-0R92) Kit 016-1R24 RAFT 24-well Bundle Kit, contains (016-0R94 & 016-1R32) Kit 016-1R25 RAFT 24-well Insert Bundle Kit, contains (016-0R94 & 016-1R33) Kit 016-1R16 RAFT Small Kit, 12 reactions, reagent and 24-well plate absorbers Kit 016-1R32 RAFT Absorbers, for 24-well Plate R33 RAFT Insert Absorbers, for 24-well Plate, 48 inserts 016-0R92 RAFT Plate Kit, 4 96-well clear plate and 4 96-well absorber Kit 016-0R94 RAFT Reagent Kit, Reagent Kit for 3D Cell Cultures Kit Clonetics Primary Animal Neurons R-G-535 Rat Microglia 2 million cells M-Cx-400 Mouse CD1 Brain Cortex Neurons 4 million cells M-Cp-402 Mouse CD1 Brain Striatum Neurons 4 million cells M-Cx- 300 Mouse C57 Brain Cortex Neurons 4 million cells M-Cp-302 Mouse C57 Brain Striatum Neurons 4 million cells M-Hi-401 Mouse CD1 Brain Hippocampus Neurons 1 million cells R-Cx-500 Rat Brain Cortex Neurons 4 million cells R-Hi-501 Rat Brain Hippocampus Neurons 1 million cells R-Hth-507 Rat Brain Hypothalamus Neurons 2 million cells R-Cp-502 Rat Brain Striatum Neurons 4 million cells R-Drg-505 Rat Dorsal Root Ganglion Neurons 0.2 million cells R-eDRG-515 Rat Dorsal Root Ganglion Neurons (embryonic) 1 million cells R-Cb-503 Rat Cerebellar Neurons 4 million cells R-Ret-508 Rat Retinal Cells 0.2 million cells Clonetics Primary Animal and Human Astrocytes CC-2565 NHA Normal Human Astrocytes 1 million cells M-AsM-430 Mouse CD1 Brain Mixed Astrocytes 1 million cells M-AsM-330 Mouse C57 Brain Mixed Astrocytes 1 million cells R-CxAs-520 Rat Brain Cortex Astrocytes 1 million cells R-HiAs-521 Rat Brain Hippocampus Astrocytes 1 million cells R-CpAs-522 Rat Brain Striatum Astrocytes 1 million cells R-AsM-530 Rat Brain Cx-Hi-Cp Mix Astrocytes 1 million cells Poietics Neural Progenitor Cells PT-2599 NHNP Normal Human Neural Progenitor Cells 1.2 million cells 16

17 Ordering Information Cat. No. Description Size Clonetics /Poietics Primary Neural Cell Growth Media CC-4461 CC-4512 PNGM Primary Neuron Growth Medium BulletKit Includes Basal Medium and SingleQuots Kit PNGM -A Primary Neuron Growth Medium Adult BulletKit Includes Basal Medium and SingleQuots Kit CC-3256 PNBM Basal Medium 500 ml CC-4462 PNGM SingleQuots Supplement and Growth Factors CC-4511 PNGM -A Primary Neuron Growth Medium Adult SingleQuots Kit Kit CC-3186 AGM BulletKit Kit Includes Basal Medium and SingleQuots Kit Kit CC-3187l ABM Basal Medium 500 ml CC-4123 AGM SingleQuots Supplements and Growth Factors CC-3209 NPMM Neural Progenitor Maintenance Medium BulletKit Kit CC-3229 NPDM Neural Progenitor Differentiation Medium BulletKit Kit CC-3210 NPBM Neural Progenitor Basal Medium 200 ml CC-4241 Neural Progenitor Maintenance Medium SingleQuots Kit (contains hegf and hfgf) CC-4242 Neural Progenitor Supplement SingleQuots Kit (contains NSF-1 and GA) Kit Nucleofector Devices AAB-1001 Nucleofector 2b Device AAF-1001B 4D-Nucleofector Core Unit AAF-1001X 4D-Nucleofector X Unit AAF-1001Y 4D-Nucleofector Y Unit AAM-1001S 96-well Shuttle Device Kits for 4D-Nucleofector Device V4XP-3012 P3 Primary Cell 4D-Nucleofector X Kit L (for neurons and glial cells) 12 rxn (100 μl Nucleocuvette ) V4XP-3024 P3 Primary Cell 4D-Nucleofector X Kit L (for neurons and glial cells) 24 rxn (100 μl Nucleocuvette ) V4XP-3032 P3 Primary Cell 4D-Nucleofector X Kit S (for neurons and glial cells) 32 rxn (20 μl Nucleocuvette ; 16-well) V4YP-1A24 Basic Neuron 4D-Nucleofector Y AD Kit (adherent) 24 rxn (Dipping Electrode) V4XP-9096 Primary Cell Optimization 4D-Nucleofector X Kit (for neural stem cells) 96 rxn (20 μl Nucleocuvette ; 16-well) Kit Kit Kit Continued on Next Page 17

18 BioResearch Neurobiology Research Tools Ordering Information Cat. No. Description Size Kits for 96-well Shuttle Device V4SP-3096 P3 Primary Cell 96-well Nucleofector Kit (for neurons and glial cells) 96 rxn (20 μl Nucleocuvette ; 96-well) V4SP-3960 P3 Primary Cell 96-well Nucleofector Kit (for neurons and glial cells) 960 rxn (20 μl Nucleocuvette ; 96-well) V4SP-9096 Primary Cell Optimization 96-well Nucleofector Kit (for neural stem cells) 192 rxn (20 μl Nucleocuvette ; 96-well) Kits for Nucleofector II/2b Device VAPG-1001 Mouse Neuron Nucleofector Kit 10 rxn (100 μl aluminum cuvette) VPG-1001 Mouse Neuron Nucleofector Kit 25 rxn (100 μl aluminum cuvette) VVPG-1001 Mouse Neuron Nucleofector Kit 4 25 rxn (100 μl aluminum cuvette) VAPG-1003 Rat Neuron Nucleofector Kit 10 rxn (100 μl aluminum cuvette) VPG-1003 Rat Neuron Nucleofector Kit 25 rxn (100 μl aluminum cuvette) VVPG-1003 Rat Neuron Nucleofector Kit 4 25 rxn (100 μl aluminum cuvette) VAPI-1003 Basic Nucleofector Kit for Primary Mammalian Neurons 10 rxn (100 μl aluminum cuvette) VPI-1003 Basic Nucleofector Kit for Primary Mammalian Neurons 25 rxn (100 μl aluminum cuvette) VVPI-1003 Basic Nucleofector Kit for Primary Mammalian Neurons 4 25 rxn (100 μl aluminum cuvette) VAPI-1006 Basic Nucleofector Kit for Primary Mammalian Glial Cells 10 rxn (100 μl aluminum cuvette) VPI-1006 Basic Nucleofector Kit for Primary Mammalian Glial Cells 25 rxn (100 μl aluminum cuvette) VVPI-1006 Basic Nucleofector Kit for Primary Mammalian Glial Cells 4 25 rxn (100 μl aluminum cuvette) VAPG-1004 Mouse Neural Stem Cell (NSC) Nucleofector Kit 10 rxn (100 μl aluminum cuvette) VPG-1004 Mouse Neural Stem Cell (NSC) Nucleofector Kit 25 rxn (100 μl aluminum cuvette) VVPG-1004 Mouse Neural Stem Cell (NSC) Nucleofector Kit 4 25 rxn (100 μl aluminum cuvette) VAPG-1005 Rat Neural Stem Cell (NSC) Nucleofector Kit 10 rxn (100 μl aluminum cuvette) VPG-1005 Rat Neural Stem Cell (NSC) Nucleofector Kit 25 rxn (100 μl aluminum cuvette) VVPG-1005 Rat Neural Stem Cell (NSC) Nucleofector Kit 4 25 rxn (100 μl aluminum cuvette) BioAssay Kits for Cell Proliferation and Cytotoxicity LT ViaLight Plus Cell Proliferation and Cytotoxity BioAssay Kit 500 tests LT ,000 tests LT ,000 tests LT tests (with 5 white TC plates) LT ToxiLight Non-destructive Cytotoxity BioAssay Kit 500 tests LT ,000 tests LT tests (with 5 white TC plates) 18

19 Ordering Information PAGEr Gels and Accessories Cat. No. Cat. No. Cat. No. Cat. No. Cat. No. Cat. No. Gel Concentration/Separation Range Cassette Size (cm) 2D well 10 well 12 well 16 well 17 well well 412 % gradient kda 420 % gradient 5200 kda 816 % gradient kda 1020 % gradient 5150 kda 7.5 % kda 10 % kda 12 % kda 15 % 1050 kda For PAGEr Gel formats and accessories. 19

20 Contact Information North America Customer Service: (toll free) Scientific Support: (toll free) Europe Customer Service: Scientific Support: International Contact your local Lonza distributor Customer Service: Fax: International Offices Australia Belgium Brazil China France (toll free) Germany (toll free) India Japan Luxemburg Singapore The Netherlands (toll free) United Kingdom (toll free) Lonza Cologne GmbH Cologne Germany For research use only. Not for use in diagnostic procedures. Neurobasal and B27 Serum Supplement are registered trademarks of Life Technologies. The Nucleofector Technology is covered by patent and/or patent pending rights owned by the Lonza Group Ltd or its affiliates. All trademarks belong to Lonza or its affiliates or to their respective third party owners.the information contained herein is believed to be correct and corresponds to the latest state of scientific and technical knowledge. However, no warranty is made, either expressed or implied, regarding its accuracy or the results to be obtained from the use of such information and no warranty is expressed or implied concerning the use of these products. The buyer assumes all risks of use and/or handling. Any user must make his own determination and satisfy himself that the products supplied by Lonza Group Ltd or its affiliates and the information and recommendations given by Lonza Group Ltd or its affiliates are (i) suitable for intended process or purpose, (ii) in compliance with environmental, health and safety regulations, and (iii) will not infringe any third party s intellectual property rights Lonza Cologne GmbH. All rights reserved. CD-BR014 CGU 05/16