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N21-MAX Media Supplement (50X)
R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
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N21-MAX Reduces Spontaneous Stress Rod Formation. E18 rat hippocampal neuron cultures were plated on poly-D-lysine coated coverslips and grown for 5 days in media supplement with either N21-MAX (Catalog # AR008) or another commercial neural media supplement (Competitor). Cells were fixed and immunostained for rod formation, and indicator of cellular stress, using an antibody against cofilin (% of Neurons with Rods). A lower percentage of hippocampal neurons that were cultured in N21-MAX had spontaneous cofilin rod formation (4.5%) compared to neurons cultured in competitor media (16.8%). Data courtesy of the laboratory of Dr. James Bamburg at Colorado State University. |
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Improved Resolution of Rod Induction in Neurons Cultured in N21-MAX. E18 rat hippocampal neuron cultures were plated on poly-D-lysine coated coverslips and grown for 5 days in N21-MAX (Catalog # AR008). Neurons were then treated with rod-inducing agents or maintained in N21-MAX alone (Untreated). Neurons were fixed and immunostained for rod formation using an antibody against cofilin (% of Neurons with Rods). The low background of rod formation in untreated cultures enabled detection of mild rod induction using Synthetic Amyloid beta, Amyloid beta dimer/trimer, and TNF-alpha. As a positive control, treatment with an ATP depletion solution or glutamate resulted in robust rod induction. Data courtesy of the laboratory of Dr. James Bamburg at Colorado State University. |
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N21-MAX Media Supplement Enhances Synaptic Development. E18 rat hippocampal neurons were grown for 19 days in vitro in media supplemented with either N21-MAX Media Supplement or the neural media supplement from the most widely-used competitor. Cells were incubated with the synaptic vesicle dye, SynaptoRed™C2 (10 µM; Catalog # 5118), for 1 minute prior to depolarization with 50 mM KCl. Cells were imaged for SynaptoRed™C2-positive synaptic puncta using the Operetta CLS™High Content Analysis System. (A) Quantification of SynaptoRed™C2-positive puncta shows that neurons grown in N21-MAX have an increased number of synaptic puncta compared to the competitor media. (B) Quantification of dye intensity shows that neurons grown in N21-MAX have more robust synaptic activity than neurons cultured in the competitor media. (C) Representative images of SynaptoRed™-C2 staining in neurons cultured in N21-MAX or competitor media. (D) Representative images showing the quantification of synaptic puncta in neurons cultured in N21-MAX or competitor media. Colored circles indicate a SynaptoRed™-positive synaptic puncta. |
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Synaptotagmin and CAM Kinase II Expression in Neurons Cultured in Media Supplemented with N21-MAX. Synaptotagmin and CAM Kinase II Expression in Neurons Cultured in Media Supplemented with N21-MAX.E18 rat hippocampal neurons were grown for 21 daysin vitroin media supplemented with N21-MAX Media Supplement (Catalog # AR008). Cells were stained with either (A) a Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364), followed by the NorthernLights™(NL)557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; yellow), or (B) a Mouse Anti-Human CaM Kinase II alpha Monoclonal Antibody (Catalog # MAB5584), followed by the NL557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007; green). The nuclei were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips. |
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Increased Synaptic Puncta and Neurite Outgrowth of Primary Neurons Cultured in N21-MAX. Increased Synaptic Puncta and Neurite Outgrowth of Primary Neurons Cultured in N21-MAX.E18 rat hippocampal neurons were grown for 21 daysin vitroin media supplemented with either N21-MAX Media Supplement (Catalog # AR008) or the neural media supplement from the most widely-used competitor. Staining for Synaptotagmin (yellow) showed more robust synaptic puncta and increased neurite outgrowth in neurons cultured in N21-MAX compared to those cultured in competitor media. Cells were staineed with a Mouse Anti-Rat Synaptotagmin-1 Monoclonal Antibody (Catalog # MAB4364) followed by the NorthernLights™(NL)557-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL007). Nuclei were counterstained with DAPI (blue). |
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N21-MAX Media Supplement Increases Efficiency of Pancreatic Cell Differentiation. N21-MAX Media Supplement Increases Efficiency of Pancreatic Cell Differentiation.Differentiation of PDX-1+pancreatic cells from induced pluripotent stem cells (iPS2) was performed with base media containing either the N21-MAX Media Supplement (green) or the competitor equivalent (red).A) Compared to competitor media, N21-MAX improved pancreatic cell differentiation as observed by flow cytometry.B) Bar graph quantifying PDX-1+cells in N21-MAX and the competitor media. |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, completed N21-MAX-supplemented 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 N21-MAX Media Supplement (Catalog # AR008):
- Albumin (bovine)
- L-Carnitine
- Catalase
- Corticosterone
- Ethanolamine
- Glutathione
- Galactose
- Holo-Transferrin
- Insulin
- Linoleic Acid
- Linolenic Acid
- Lipoic Acid
- Progesterone
- Putrescine
- Retinyl acetate
- Retinol
- Selenite
- Superoxide dismutase
- Triiodo-L-thyronine
- D,L-alpha-Tocopherol
- D,L-alpha-Tocopherol acetate
Other Supplies Required
Reagents
- Basal media (e.g., Neurobasal Media from Invitrogen®, Catalog # 21103-029 or equivalent)
- L-Glutamine
Materials
- Serological pipettes
- Pipettes and pipette tips
Equipment
- 2 °C to 8 °C refrigerator
Protocol Overview
Dilute 50-fold with a basal media (e.g., Neurobasal Media from Invitrogen, Catalog # 21103-029) and supplement with 0.5 mM L-glutamine 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 for N21-MAX Media Supplement (50X) (17)
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Citations are publications that use Bio-Techne products. Selected citations for N21-MAX Media Supplement (50X) include:
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Species: Rat
Sample Types: Whole Cells
Applications: BioassayM Jhanji et al. (2022), Cis- and trans-resveratrol have opposite effects on histone serine-ADP-ribosylation and tyrosine induced neurodegeneration Nature Communications, 13(1):3244.
PMID: 35688816 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayM Mehrabian et al. (2022), Cardiac glycoside-mediated turnover of Na, K-ATPases as a rational approach to reducing cell surface levels of the cellular prion protein PLoS ONE, 17(7):e0270915.
PMID: 35776750 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayD Liu et al. (2022), Impact of schizophrenia GWAS loci converge onto distinct pathways in cortical interneurons vs glutamatergic neurons during development Molecular Psychiatry, 0(0).
PMID: 35701597 -
Species: Human
Sample Types: Whole Cells
Applications: Cell culture supernatesYX Xie et al. (2022), Lysosomal exocytosis releases pathogenic alpha-synuclein species from neurons in synucleinopathy models Nature Communications, 13(1):4918.
PMID: 35995799 -
Species: Rat
Sample Types: Whole Cells
Applications: BioassayM Lam et al. (2022), CNS myelination requires VAMP2/3-mediated membrane expansion in oligodendrocytes Nature Communications, 13(1):5583.
PMID: 36151203 -
Species: Human
Sample Types: Whole Cells
Applications: DifferentiationTM Triolo et al. (2022), Stem-Cell-Derived beta-Like Cells with a Functional PTPN2 Knockout Display Increased Immunogenicity Cells, 11(23).
PMID: 36497105 -
Species: Mouse
Sample Types: Whole Cells
Applications: BioassayB Sun et al. (2021), Regulation of host and virus genes by neuronal miR-138 favours herpes simplex virus 1 latency Nature Microbiology, 0(0).
PMID: 33558653 -
Species: Mouse
Sample Types: Whole Cells
Applications: BioassayD Williams et al. (2021), The cellular prion protein interacts with and promotes the activity of Na,K-ATPases PLoS ONE, 16(11):e0258682.
PMID: 34847154 -
Species: Mouse
Sample Types: Whole Cells
Applications: BioassayLK Smith et al. (2021), Direct interaction of HIV gp120 with neuronal CXCR4 and CCR5 receptors induces cofilin-actin rod pathology via a cellular prion protein- and NOX-dependent mechanism PLoS ONE, 16(3):e0248309.
PMID: 33705493 -
Species: Mouse
Sample Types: Whole Cells
Applications: BioassayY Yang et al. (2020), Silencing of AURKA augments the antitumor efficacy of the AURKA inhibitor MLN8237 on neuroblastoma cells Cancer Cell Int., 20(0):9.
PMID: 31920463 -
Species: Mouse
Sample Types: Whole Cells
Applications: Cell CultureH Jiang et al. (2020), Live Neuron High-Content Screening Reveals Synaptotoxic Activity in Alzheimer Mouse Model Homogenates Sci Rep, 10(1):3412.
PMID: 32098978 -
Species: Mouse
Sample Types: Whole Cells
Applications: BioassayY Rong et al. (2019), Neural stem cell small extracellular vesicle-based delivery of 14-3-3t reduces apoptosis and neuroinflammation following traumatic spinal cord injury by enhancing autophagy by targeting Beclin-1 Aging (Albany NY), 11(18):7723-7745.
PMID: 31563124 -
Species: Human
Sample Types: Whole Cells
X Wang et al. (2019), Tau interactome analyses in CRISPR-Cas9 engineered neuronal cells reveal ATPase-dependent binding of wild-type but not P301L Tau to non-muscle myosins Sci Rep, 9(1):16238.
PMID: 31700063 -
Species: Human
Sample Types: Whole Cells
M Seo et al. (2018), Stemness-Attenuating miR-503-3p as a Paracrine Factor to Regulate Growth of Cancer Stem Cells Stem Cells Int, 2018(0):4851949.
PMID: 29849663 -
Species: Human
Sample Types: Whole Cells
PK Sahoo et al. (2018), Axonal G3BP1 stress granule protein limits axonal mRNA translation and nerve regeneration Nat Commun, 9(1):3358.
PMID: 30135423 -
Species: Rat
Sample Types: Whole Cells
Lariosa-Willingham K et al. (2016), Development of a central nervous system axonal myelination assay for high throughput screening. BMC Neurosci, 17(0):16.
PMID: 27103572 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayYang Y et al. (0), Silencing of AURKA augments the antitumor efficacy of the AURKA inhibitor MLN8237 on neuroblastoma cells. Cancer Cell In20(0):9.
PMID: 31920463
There are no citations that match your criteria.
Customer Reviews for N21-MAX Media Supplement (50X) (6)
4.3 out of 5
6 customer ratings
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Verified Customer | Posted 05/19/2021
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Verified Customer | Posted 12/20/2018The area, intensity, and density of PSD-95 signal are comparable to the culture grown in B27.
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Verified Customer | Posted 04/04/2018
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Verified Customer | Posted 10/30/2017Adult human stem cells showed fast growth and took less time to differentiate into neurons. Data is communicated so cannot upload image.
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Verified Customer | Posted 10/30/2017I have previously used Gibco B27®, and Gem21 NeuroPlex™ for culturing of neural cells and neuronal lines. I use typical for co- culture of neurons with astrocytes and microglia. I tried your sample N21 max from R&D and it worked very well with my neuronal cells co cultures. All of these Gibco B27®, and Gem21 NeuroPlex™ and N21 max are Supplied as a 50X concentrate and are list priced at $79 for 10 ml. I would use them all interchangeably as they all worked equally well in my co culture system.
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Verified Customer | Posted 12/06/2016For stem cell culture media supplementation.
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FAQs for N21-MAX Media Supplement (50X)
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What advantages does the N21-MAX Media Supplement (Catalog # AR008) have over other neuronal stem cell supplements?
We have performed comparison testing in-house and have found that in long term cultures (21 days), we observe better synapse marker expression.
Additionally, one major formulation difference is that our N21 contains holo-transferrin while other supplements typically contain apo-transferrin. It is reported in the literature that holo-transferrin is preferred for neuronal cell culture. We have also tested several BSA sources to determine which is optimal for neuronal cultures.
