N-2 MAX Media Supplement (100X) Best Seller
R&D Systems, part of Bio-Techne | Catalog # AR009
Key Product Details
Summary for N-2 MAX Media Supplement (100X)
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 °C in a manual defrost freezer. Do not use past the expiration date.
Precautions
This product contains human transferrin. This transferrin was purified from donor plasma and 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.
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.
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.
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Neural Progenitor Cells Expanded with N-2 MAX Media Supplement Express Nestin and SOX2. Neural Progenitor Cells Expanded with N-2 MAX 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 IgG2AIsotype 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. |
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Lot Consistency of N-2 MAX Media Supplement. Rat cortical stem cells (RCSC) were cultured over multiple passages using N-2 MAX and evaluated via flow cytometry for the maintenance of stem cell markers, SOX2 and Nestin. Quantification of flow cytometry data demonstrates a high purity of (A) SOX2 or (B) Nestin positive RCSCs across multiple lots of N-2 MAX Supplement. C) Average SOX2 and Nestin-positive RCSCs, demonstrate the consistent lot-to-lot performance of N-2 MAX. |
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Verification of Neural Progenitor Cell Multipotency Following Expansion with N-2 MAX Media Supplement. 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 daysin vitroin 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. |
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N-2 MAX Media Supplement is Comparable to GMP N-2 MAX. Rat Cortical Stem Cells (Catalog # NSC001) were cultured for 14 days in media supplemented with 20 ng/mL Recombinant Human FBF basic (Catalog # 233-FB) and either N-2 MAX Media Supplement or GMP N-2 MAX Media Supplement (Catalog # AR016). Cells were harvested and stained with PE-conjugated Mouse Anti-Human Nestin Monoclonal Antibody (Catalog # IC1259) or PE-conjugated Anti-Human/Mouse SOX2 Monoclonal Antibody (Catalog # IC2018). Under these conditions, similar yields of Nestin and SOX2 positive cells were observed. |
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 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)
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
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.