N21-MAX Media Supplement (50X)

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.

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.

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.

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.

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).

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.