Kit Summary

For culturing neurons, neural progenitors, and stem cells.

Key Benefits

  • Optimized for neural and stem cell cultures
  • Recombinant Human Insulin ensures low experimental variability
  • Consistent, chemically-defined, and serum-free formulation
  • Ideal for NPC derivation, maintenance, and differentiation
  • Enhances performance of stem cell differentiation protocols
 

Why Culture Neurons under Fully Defined Conditions?

Serum-free, defined media are routinely used as an alternative to standard serum-containing media in order to reduce unwanted experimental variability.

Uncontrolled variables commonly associated with serum-supplemented media, such as indeterminate levels of vitamins, hormones, and growth factors are eliminated, and the potential for contamination by infectious agents is reduced when cells are cultured under defined conditions. In addition, serum-free media offers researchers the ability to specifically design the culture media for a particular experimental question.

The N-2 MAX Media Supplement:

  • Contains high quality factors to support reproducible and efficient NPC expansion.
  • Is fully defined to reduce unwanted experimental variability.
  • Has been developed and optimized using neural progenitor cells.

Contents

Supplied as a 100X concentrate in water, this media supplement contains the following high quality factors to support neural cell culture:

 
Component Amount (mg/mL)
Recombinant Human Insulin 2,500 µg/mL
Human Transferrin 10,000 µg/mL
Putrescine 1,611 µg/mL
Selenite 0.52 µg/mL
Progesterone 0.63 µg/mL

Supplied in a volume sufficient to supplement 500 mL of media at the recommended concentration.

 

Stability and Storage

Store in the dark at < -20 °C in a manual defrost freezer. Do not use past the expiration date.

Precautions

This product contains human transferrin. This transferrin was tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen. As no testing can offer complete assurance of freedom from infectious reagents, these reagents should be handled as if capable of transmitting infection.

Limitations

  • FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
  • The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
  • This reagent should not be used beyond the expiration date indicated on the label.

 

Data Examples
Neural Progenitor Cells Expanded with N-2 Plus Media Supplement Express Nestin and SOX2.
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Neural Progenitor Cells Expanded with N-2 Plus Media Supplement Express Nestin and SOX2. Rat Cortical Stem Cells (Catalog # NSC001) were cultured for 7 days in media supplemented with 1X N-2 MAX Media Supplement (Catalog # AR009) and 20 ng/mL of Recombinant Human FGF basic (Catalog # 233-FB). The cells were stained with a PE-conjugated Mouse Anti-Human Nestin Monoclonal Antibody (Catalog # IC1259P; red histogram), a PE-conjugated Mouse Anti-Human/Mouse SOX2 Monoclonal Antibody (Catalog # IC2018P; green histogram), or a PE-conjugated Mouse IgG2A Isotype Control (Catalog # IC003P; open histogram). Under these conditions, cells were shown to express high levels of Nestin and SOX2, two established markers of neural multipotency.

Verification of Neural Progenitor Cell Multipotency Following Expansion with N-2 Plus Media Supplement
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Verification of Neural Progenitor Cell Multipotency Following Expansion with N-2 MAX Media Supplement. Rat Cortical Stem Cells (Catalog # NSC001) were grown and differentiated for 7 days in vitro in media supplemented with N-2 MAX Media Supplement (Catalog # AR009). Markers of lineage differentiation were detected using a Mouse Anti-Neuron-specific beta-III Tubulin Monoclonal (clone TuJ-1) Antibody (Catalog # MAB1195), followed by a NorthernLights (NL)557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL007; red), a Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594), followed by a NL557-conjugated Donkey Anti-Sheep Secondary Antibody (Catalog # NL010; red), and a Mouse Anti-Human/Mouse/Rat/Chicken Oligodendrocyte Marker O4 Monoclonal Antibody (Catalog # MAB1326), followed by a NL 557-conjugated Goat Anti-Mouse Secondary Antibody (Catalog # NL019; red). The nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Preparation and Storage
  • Stability & Storage
    Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Background: Neural Stem Cells

Neural stem cells provide an excellent model for research focused on neural development and neurological disorders. R&D Systems offers ready-to-use primary cortical stem cells isolated from E14.5 Sprague-Dawley rats. In addition, primary mouse cortical stem cells isolated from E14.5 CD-1 mice are available. Every lot of R&D Systems Cortical Stem Cells is validated for a high level of Nestin expression and the capacity for multi-lineage differentiation into astrocytes, neurons, and oligodendrocytes. Our cortical stem cells are tested to ensure highest quality and lot to lot consistency. Both rat and mouse Cortical Stem Cells can be optimally expanded as monolayers or neurospheres.

To complement the use of primary neural stem cells, we offer a range of supportive products, including culture media which is specifically optimized for use with neural stem cells. We offer kits to promote the in vitro proliferation of neural precursors, and kits to differentiate neural stem cells into dopaminergic neurons or oligodendrocytes. In addition, kits are available which contain panels of antibodies designed to monitor the differentiation and identification of neural precursors, astrocytes, neurons, and oligodendrocytes.

  • Alternate Names:
    Neural Stem Cells
Assay Procedure

Refer to the product datasheet for complete product details.

 

Briefly, completed N-2 MAX-supplemented neural cell medium is prepared using the following procedure:

  • Dilute the media supplement in basal media
  • Store completed media
  • Use within 2 weeks
 

 

Reagents Provided

Reagents provided in the N-2 MAX Media Supplement (Catalog # AR009):

  • Recombinant Human Insulin (2,500 µg/mL)
  • Human Transferrin (10,000 µg/mL)
  • Putrescine (1,611 µg/mL)
  • Selenite (0.52 µg/mL)
  • Progesterone (0.63 µg/mL)

 

Other Supplies Required

Reagents

  • DMEM/F-12 (Invitrogen®, Catalog # 12500-062) or a basal media (e.g., Neurobasal Media from Invitrogen, Catalog # 21103-029)
  • Glucose
  • Glutamine
  • NaHCO3
  • Penicillin-Streptomycin (100X)
  • Deionized or distilled water

Materials

  • Serological pipettes
  • Pipettes and pipette tips
  • 2 µm filter unit

Equipment

  • 2 °C to 8 °C refrigerator
 

 

Protocol Overview

Option 1: Mix the following components with deionized or distilled water to make 500 mL of medium.

Component Amount (mg/mL)
DMEM/F-12 6 g
Glucose 0.775 g
Glutamine 0.0365 g
NaHCO3 0.854 g
N-2 MAX Media Supplement 5 mL

 

Adjust the pH to 7.2. Filter the solution (2 µm filter unit), and add 5 mL of 100X sterile Penicillin-Streptomycin solution. The medium may be stored in the dark at 2 °C to 8 °C for up to 2 weeks.

Option 2: Dilute 100-fold with a basal media (e.g., Neurobasal Media from Invitrogen, Catalog # 21103-029) before use. The medium may be stored in the dark at 2 °C to 8 °C for up to 2 weeks.

Invitrogen is a registered trademark of Invitrogen Corp.

Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
Filter your results:

Species
Applications
Sample Type
  1. Isolation and characterization of string-forming female germline stem cells from ovaries of neonatal mice
    Authors: J Liu, D Shang, Y Xiao, P Zhong, H Cheng, R Zhou
    J. Biol. Chem., 2017;0(0):. 2017
  2. Isolation, characterization and propagation of mitotically active germ cells from adult mouse and human ovaries.
    Authors: Woods D, Tilly J
    Nat Protoc, 2013;8(5):966-88. 2013
  3. Transcription factor-mediated reprogramming of fibroblasts to expandable, myelinogenic oligodendrocyte progenitor cells.
    Authors: Najm, Fadi J, Lager, Angela M, Zaremba, Anita, Wyatt, Krysta, Caprariello, Andrew V, Factor, Daniel C, Karl, Robert T, Maeda, Tadao, Miller, Robert H, Tesar, Paul J
    Nat Biotechnol, 2013;31(5):426-33. 2013
  4. The helicase HAGE expressed by malignant melanoma-initiating cells is required for tumor cell proliferation in vivo.
    Authors: Linley AJ, Mathieu MG, Miles AK
    J. Biol. Chem., 2012;287(17):13633-43. 2012
  5. Modeling the Mutational and Phenotypic Landscapes of Pelizaeus-Merzbacher Disease with Human iPSC-Derived Oligodendrocytes.
    Authors: Nevin Z, Factor D, Karl R, Douvaras P, Laukka J, Windrem M, Goldman S, Fossati V, Hobson G, Tesar P
    Am J Hum Genet, 0;100(4):617-634. 0
  6. Neuronal differentiation of hair-follicle-bulge-derived stem cells co-cultured with mouse cochlear modiolus explants.
    Authors: Schomann T, Mezzanotte L, De Groot J, Rivolta M, Hendriks S, Frijns J, Huisman M
    PLoS ONE, 0;12(10):e0187183. 0
Mycoplasma Detection
Description Application Cat# Citations Images  

MycoProbe Mycoplasma Detection Kit

CUL001B 29
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Average Rating: 4.5 (Based on 6 reviews)

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Images Ratings Applications Species Reviewed By Date Details
 N-2 MAX Media Supplement (100X) AR009
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Excellent
  Anonymous 11/01/2017
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Image Details
 N-2 MAX Media Supplement (100X) AR009
: N-2 MAX Media Supplement (100X) [AR009]

Other Experimental Details

Other Experimental DetailsIPSCs cultured on a polymer supported plate for differentiation
 N-2 MAX Media Supplement (100X) AR009
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Very Good
  Anonymous 05/30/2017
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 N-2 MAX Media Supplement (100X) AR009
: N-2 MAX Media Supplement (100X) [AR009]

Other Experimental Details

Other Experimental Details I have previously used N2 Neuroplex supplement from Gemimi Bio (Cat # 400-163) to culture neurons and this sample from R&D Systems work equally well. We will be switching to N-2 MAX.
  Very Good
  Anonymous 12/06/2016
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Other Experimental Details

Other Experimental DetailsN2 Max is a component of our base media for differentiating mouse and human stem cells to neural progenitor cells for modeling neural development and disease.
  Excellent
  Anonymous 12/06/2016
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Other Experimental Details

Other Experimental DetailsI use this product to grow Neural Progenitor Cells (NPC). This product is excellent. The cells grow very well in this media.
  Very Good
  Anonymous 12/06/2016
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Other Experimental Details

Other Experimental DetailsWe use in our media for neural stem cell expansion.
  Excellent
  Anonymous 12/06/2016
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Other Experimental Details

Other Experimental DetailsN2 max supplement was used throughout the whole process of generation, culture, and maintenance of human and mouse iPS-derived dopaminergic neurons. The product performed well and better than one produced by another company.

FAQs

  1. What is the difference between N-2 Plus Media Supplement (Catalog # AR003) and N-2 MAX Media Supplement (Catalog # AR009)?

    • The N-2 Plus Media Supplement (Catalog # AR003) contains Bovine Insulin whereas N-2 MAX Media Supplement (Catalog # AR009) contains Recombinant Human Insulin. All other media components and concentrations in N-2 Plus and N-2 MAX are the same. These two products perform comparably in side-by-side testing. Due to a limited supply of bovine insulin, the products are priced differently.

  2. What is the difference between N-2 Plus Media Supplement (Catalog # AR003) and N-2 MAX Media Supplement (Catalog # AR009)?

    • The N-2 Plus Media Supplement (Catalog # AR003) contains Bovine Insulin whereas N-2 MAX Media Supplement (Catalog # AR009) contains Recombinant Human Insulin. All other media components and concentrations in N-2 Plus and N-2 MAX are the same. These two products perform comparably in side-by-side testing. Due to a limited supply of bovine insulin, the products are priced differently.

  3. Does N-2 MAX Media Supplement (Catalog # AR009) contain any components that are calcium salts?

    • No.
  4. Does N-2 MAX Media Supplement (Catalog # AR009) contain any components that are calcium salts?

    • No.
  5. Does N-2 MAX Media Supplement (Catalog # AR009) contain any proteins that are produced using Baculovirus?

    • No.
  6. Does N-2 MAX Media Supplement (Catalog # AR009) contain any proteins that are produced using Baculovirus?

    • No.
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