Neurobiology Research Tools for Biologically Relevant Results

Transcription

1 BioResearch Neurobiology Research Tools for Biologically Relevant Results Os nos ad tat. Igna faccum eroses iriusti onulla feuisisit adio con veliquamcore magna alis nonulla non er irilla am, quiscil dolobor eu faccum quisisim quis nullaor rem iril er summy nulpute tinos ipisci te facilit nulla facilis molore. 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 mrna, protein expression, cell proliferation, or cytotoxicity." SOLUTION: Choose from our variety of neurology-specific assay tools, including qpcr arrays, ready-to-use protein gels, and bioassay kits to measure cell proliferation and cytotoxicity. Page 14 Cell Analysis " I need human model systems to avoid species extrapolation in my drug discovery process." SOLUTION: Now you can access assay-ready embryonic stem cell-derived pure human motor neurons. 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 Cell Analysis Drug Discovery 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 Amaxa 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 StellARray qpcr Arrays for mrna expression profiling: Alzheimer's (mouse or human) Autism (mouse or human) Huntington's (mouse or human) Mood disorder (mouse or human) Neurodegeneration (mouse or human) Parkinson's (mouse or human) Protein Expression Analysis: PAGEr Precast Protein Gels for Western blotting Assay-ready human embryonic stem cell (ESC) derived neurons: NeuroPlate Human ESC Neuronal Progenitors MotorPlate Standard Human ESC Motor Neuron Progenitors MotorPlate Mature Human ESC Motor Neuron Progenitors 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 in your freezer and thaw them as needed. " 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. Cryopreserved Clonetics Primary Neurons 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) Recommended Media Hippocampal neurons n n PNGM Cortical neurons n n PNGM Striatum neurons n n PNGM Dorsal root ganglia (embryonic or neonatal) n PNGM Cerebellar granule cells n PNGM -A Hypothalamus neurons n PNGM Retinal neurons n PNGM Applications Transfection Evaluating electrophysiological properties, neurotransmitters, and receptor function Drug screening Research into ion channels, intracellular transport, and neurotoxicity Rat cortical neuronal cells were thawed, cultured 14 days, and immunofluorescently stained with anti-pgp 9.5 and anti ß-tubulin. Clonetics PNGM Primary Neuron Growth Media 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 Competitor 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. 5

6 BioResearch Neurobiology Research Tools Cryopreserved Clonetics Primary Astrocytes Improve your workflow with cryopreserved, high-quality human, and animal glial cells. There s 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) Hippocampal astrocytes n Cortical astrocytes n Striatum astrocytes n Cx-Hi-Cp mixed Astrocytes n Cx-Hi-Striatum mixed Astrocytes n Astrocytes n Human Indirect immunofluorescence staining of Clonetics Normal Human Astrocytes for glial fibrillary acidic protein (GFAP). Clonetics AGM Primary Astrocyte Growth Media 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 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 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 Cryopreserved primary Clonetics Rat Cortical and Striatal Neurons were thawed, plated in tandem, and incubated for 7 35 days in vitro. Cultures were characterized using synaptic marker antibodies, receptor and second messenger proteins, and structural marker proteins. Conclusion 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. Results 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. Amaxa Nucleofector Technology 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 " 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. 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 μg (20 µl) 1 5 μg (100 µl) μg μg 1 5 μg sirna amount/sample (concentration 2 nm 2 µm) pmol (20 µl) pmol (100 µl) pmol pmol pmol Adherent Nucleofection n n Compatibility with n 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 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 55-65% 60-70% P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Astrocytes, rat 60-70% 70-80% P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Dorsal root ganglia (DRG), chicken 25-35% P3 Primary Cell (Neuron, Basic) Chicken Neuron Dorsal root ganglia (DRG), rat 35-45% P3 Primary Cell (Neuron, Basic) Rat Neuron Neuron, cortical, rat 30-70% 45-60% P3 Primary Cell (Neuron, rat) Rat Neuron Neuron, hippocampal, chicken 40-50% P3 Primary Cell (Neuron, Basic) Chicken Neuron Neuron, hippocampal, mouse 50-60% P3 Primary Cell (Neuron, Basic) Mouse Neuron Neuron, hippocampal, rat 30-70% 45-60% P3 Primary Cell (Neuron, rat) Rat Neuron Neuron, hippocampal, rat adherent 30-50% 50-80% Basic Neuron AD (Neuron, Basic, AD) Not Applicable Neuron, cortical, rat adherent 30-60% 70-90% Basic Neuron AD (Neuron, Basic, AD) Not Applicable Neuron, cortical, mouse adherent 20-70% 70-90% Basic Neuron AD (Neuron, Basic, AD Not Applicable Oligodendrocyte, rat 40-50% 55-65% P3 Primary Cell (Neuron, Basic) Glial Cell, Basic Stem Cells Neural stem cell (NSC), mouse 75-85% Primary Cell Optimization Mouse NSC Neural stem cell (NSC), rat 40-50% Primary Cell Optimization Rat NSC Neuronal Cell Lines** AGN2a 60-95% Cell Line Optimization Cell Line R BV % 70-95% Cell Line Optimization Cell Line T C % 70-80% Cell Line Optimization Cell Line V H % 80-90% Cell Line Optimization Cell Line V IMR % 55-65% Cell Line Optimization Cell Line L Neuro2A 47-85% 80-95% Cell Line SF Cell Line V NG % 75-85% Cell Line Optimization Cell Line V PC % 75-85% Cell Line Optimization Cell Line V SH-SY5Y 60-85% 40-80% Cell Line SF Cell Line V SK-N-MC 60-95% 30-60% Cell Line Optimization Cell Line V SK-N-SH 80-90% 65-75% Cell Line Optimization Cell Line V U-87MG 35-45% 85-95% Cell Line Optimization Cell Line T Primary cells marked blue have Lonza-validated optimized protocols. Cell lines marked blue also have optimized protocols with ATCC clones. *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 System that also enables adherent Nucleofection : Achieves transfection efficiencies of up to 70% Basic Kits help pinpoint optimal conditions for primary neurons, glial cells or neural stem cells Has been tested for cryopreserved Clonetics Primary Neurons Applications 4D-Nucleofector Unit or 96-well Shuttle Add-on: Culture cells for up to 14 days in specialized Nucleocuvette AD Strips (20 μl) Transfect them at any time during this culture period Suited for analysis by light and fluorescence microscopy; absorption, luminescence, or fluorescence assays; or mrna expression 4D-Nucleofector Y Unit: Culture cells in standard 24-well plates For Nucleofection, simply insert a disposable conductive polymer 24-well Dipping Electrode Array for Nucleofection Ideal for post-transfection analysis by confocal microscopy or patch clamping Adherent Nucleofection All-in-one Process Pre-culture Preserved functionality after adherent Nucleofection of neurons. Mouse cortical neurons were cultured in 24-well plates and transfected after 6 DIV with 17.5 μg pmaxfp -Yellow-C Vector using the Basic Neuron 4D-Nucleofector Y AD Kit. Four days post Nucleofection, cells were immuno-stained for maxfp -Yellow expression (green) and synapses (red), stained for nuclei (DAPI, blue), and analyzed by fluorescence microscopy. Transfected neurons show normal synapse formation. Adherent Nucleofection of cryopreserved Clonetics Rat Hippocampal Neurons. Frozen rat hippocampal neurons were thawed and seeded into 24-well plates. After 2 days, cells were transfected with 17.5 μg pmaxgfp Vector using the Basic Neuron 4D-Nucleofector Y AD Kit. 1 day post Nucleofection, cells were analyzed by fluorescence microscopy. Up to 14 days Nucleofection or Analysis Nucleocuvette AD Strips (20 µl) Dipping Electrode Arrays Required functional 4D-Nucleofector Unit Pre- and post Nucleofection Culture X Unit or 96-well Shuttle Nucleocuvette Wells Nucleofection of cells plated on glass cover slips n High transfection efficiencies and viabilities n n Cell analysis by: transmission light or fluorescence microscopy absorption, luminescence or fluroescence assays confocal microscopy patch clamping Nucleofection of cryopreserved Clonetics Primary Animal Neurons n n n Y Unit 24-well culture plate n n n n n Adherent Nucleofection using Nucleocuvette AD Strips or 24-well Dipping Electrode Arrays. After isolation, primary neurons are directly plated into poly-d- or poly-l-lysine coated Nucleocuvette Strips (when using the Unit) or 24-well culture plates (when using the Y Unit). Cells can be cultured up to 14 days and transfected by Nucleofection at any time during this period. 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 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. 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. % Transfection Efficiency The challenge was to find a non-viral transfection method that provides high efficiency without morphological or functional changes Electroporation Lipofection Nucleofection Method To identify the best method for transfecting NPCs derived from ventral mesencephali of rat brain, the study tested electroporation, lipofection, and Nucleofection. DsRed EGFP 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 Have greater confidence in your data by using assay-ready human neurons to screen your drug candidates. Human MotorPlate Neurons Save yourself the hassle of species-to-species extrapolation by using the first commercially available source of human motor neurons in your screening campaigns: Take advantage of assay-ready human embryonic stem cellderived motor neurons Choose from different maturation stages: NeuroPlate Neural Progenitors, early stage MotorPlate Standard Motor Neurons or late stage MotorPlate Mature Motor Neurons Tested for neuronal morphology, specific markers, adherence, and density Obtain large quantities with minimal lot-to-lot variation Improve your workflow with the convenience of ready-to-use, pre-plated tools Receive optimized MotorBlast Culture Media with your shipment " I need human model systems to avoid species extrapolation in my drug discovery process." SOLUTION: Now you can access assay-ready embryonic stem cell-derived pure human motor neurons. Applications Viability, toxicity, and neuronal outgrowth assays Motor neuron function and disease research Maturation and cell cycle analysis studies Standard MotorPlate Cells stained for Neuronal Class III ß-tubulin (green) and Glial Fibrillary Acid Protein (red). Nuclei are counterstained with DAPI (blue). 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. StellARray qpcr Arrays StellARray qpcr Arrays contain 96 or 384 targeted, pre-validated qpcr assays: Choose from more than 150 pre-validated, research area-specific qpcr arrays or configure custom arrays online Use with standard qpcr equipment Perform a comprehensive range of neurobiology experiments Utilize ΔΔCt analysis, or achieve reliable normalization with StellARray Proprietary GPR Software, which automatically identifies the best normalizer amplicons based on lowest variance Human Alzheimer n n Autism n n Huntington n n Mood disorder n n Neurodegeneration n n Parkinson n n Mouse " I can t find reliable, ready-touse assay tools for mrna, protein expression, cell proliferation, or cytotoxicity." SOLUTION: Choose from our variety of neurology-specific assay tools, including qpcr arrays, ready-to-use protein gels, and bioassay kits to measure cell proliferation and 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 For more information on the StellARray Applications System and its supporting GeneSieve Search Tool or Global Pattern Recognition (GPR) Analysis Tool. SDS-PAGE Native PAGE Western blotting Applications Antibody screening Gene expression profiling Pathway analysis Comb Options and Well Volumes Microarray data validation sirna knock-down analysis Biomarker discovery 2D well 8+1 well* 12 well 550 μl sample, or 7 cm IPG strip, 12 μl marker 30 μl sample 12 μl marker 20 μl 10 well 32 μl 16 well 14 μl *Compatible with multichannel pipettes 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 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 Clonetics Primary Animal Neurons 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 PNGM Primary Neuron Growth Medium BulletKit Kit Includes Basal Medium and SingleQuots Kit CC-4512 PNGM -A Primary Neuron Growth Medium Adult BulletKit Kit 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 Kit (contains hegf and hfgf) CC-4242 Neural Progenitor Supplement SingleQuots Kit (contains NSF-1 and GA) Kit Pluripotent Stem Cell-derived Neuronal Cells FP-6020 NeuroPlate Human ESC Neuronal Progenitors 96-well plate FP-6041 NeuroPlate Human ESC Neuronal Progenitors 384-well plate FP-6011 MotorPlate Standard Human ESC Motor Neuron Progenitors 96-well plate FP-6040 MotorPlate Standard Human ESC Motor Neuron Progenitor 384-well plate FP-6046 MotorPlate Mature Human ESC Motor Neuron Progenitors 96-well plate FP-6049 MotorPlate Mature Human ESC Motor Neuron Progenitors 384-well plate Nucleofector Devices AAB-1001 Nucleofector 2b Device AAF-1001B 4D-Nucleofector Core Unit AAF-1001X 4D-Nucleofector Unit AAM-1001S 96-well Shuttle Device Kits for 4D-Nucleofector Device V4XP-3024 P3 Primary Cell 4D-Nucleofector Kit L (for neurons and glial cells) 24 rxn (100 μl Nucleocuvette ) V4XP-3032 P3 Primary Cell 4D-Nucleofector Kit S (for neurons and glial cells) 32 rxn (20 μl Nucleocuvette ; 16-well) V4XP-1A32 Basic Neuron 4D-Nucleofector AD Kit (adherent) 32 rxn (20 μl Nucleocuvette AD; 16-well) V4YP-1A24 Basic Neuron 4D-Nucleofector Y AD Kit (adherent) 24 rxn (Dipping Electrode) V4XP-9096 Primary Cell Optimization 4D-Nucleofector Kit (for neural stem cells) 96 rxn (20 μl Nucleocuvette ; 16-well) Continued on Next Page 17

18 BioResearch Neurobiology Research Tools Ordering Information Cat. No. Description Size Kits for 96-well Shuttle Device VIPI Basic Neuron 96-well Nucleofector AD Kit (adherent) 96 rxn (20 μl Nucleocuvette ; 96-well) 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 ViaLight 100% Lysis Control Set (sold separately) 10 ml (200 tests) LT ToxiLight Non-destructive Cytotoxity BioAssay Kit 500 tests LT ,000 tests LT tests (with 5 white TC plates) 18

19 Ordering Information Cat. No. Description Size CNS StellARray qpcr Assays Please Inquire Human Alzheimer s Disease StellARray Various formats Please Inquire Human Autism StellARray Various formats Please Inquire Human Huntington s Disease StellARray Various formats Please Inquire Human Mood Disorder StellARray Various formats Please Inquire Human Neurodegeneration StellARray Various formats Please Inquire Human Parkinson StellARray Various formats Please Inquire Mouse Alzheimer s Disease StellARray Various formats Please Inquire Mouse Autism StellARray Various formats Please Inquire Mouse Huntington s Disease StellARray Various formats Please Inquire Mouse Mood Disorder StellARray Various formats Please Inquire Mouse Neurodegeneration StellARray Various formats For StellARray qpcr Assay formats. 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 4 12% gradient kda 4 20% gradient kda 8 16% gradient kda 10 20% gradient kda 7.5% kda 10% kda 12% kda 15% kda For PAGEr Gel formats and accessories. 19

20 Contact Information North America Customer Service: (toll free) Scientific Support: (toll free) Europe Customer Service: order.europe@lonza.com Scientific Support: scientific.support.eu@lonza.com Online Ordering International Contact your local Lonza Distributor Customer Service: , ext scientific.support@lonza.com International Offices Australia Austria (toll free) Belgium Brazil Denmark (toll free) France (toll free) Germany (toll free) India Ireland (toll free) Italy Japan Luxemburg Poland Singapore Spain Sweden (toll free) Switzerland (toll free) The Netherlands (toll free) United Kingdom (toll free) Lonza Cologne GmbH Cologne, Germany For research use only. Not for use in diagnostic procedures. StellARray, GeneSieve, Global Pattern Recognition and GPR are trademarks of Bar Harbor Biotechnology. Neurobasal and B27 Serum Supplement are registered trademarks of Life Technologies. ATCC is a trademark of ATCC used under license. All other trademarks herein are marks of the Lonza Group or its affiliates. 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. No statement is intended or should be construed as a recommendation to infringe any existing patent Lonza Cologne GmbH. All rights reserved. BR-DseCmpgn 03/11 CD-BR014