N-2 Supplement (C007B-557136)

Aladdin's N-2 Supplement (100X) is a chemically defined 100X concentrated cell culture additive which is sterilized by filtration. N-2 Supplement promotes cell growth and survival, and is commonly used for the culture of neurons, neural stem cells, neuroblastoma cells, as well as the culture and directed differentiation of pluripotent stem cells [1-4].The composition of this product is consistent with Bottenstein's N-2 formulation [5], mainly comprising human holo transferrin, insulin recombinant full chain, progesterone, putrescine, and selenite, etc.Insulin plays an important role in the anabolism of mammalian cells, promoting cellular uptake of glucose and amino acids, lipid synthesis, transport of monovalent cations and phosphates, protein and nucleic acid synthesis, and cell proliferation. Transferrin is an important carrier of iron, reducing the toxicity of free ferric ions, oxygen free radicals, and hydrogen peroxide. Selenium in sodium selenite is a cofactor of glutathione peroxidase and some other important enzymes, and also acts as an antioxidant . Progesterone has a growth-stimulating effect on cells. Putrescine, a non-protein nitrogenous base, is a precursor of polyamines, spermidine and spermine, and may be necessary for cell division [5, 6].This product is prepared with cell culture grade ultrapure water, sterilized by filtration, and therefore can be directly used for cell culture and other applications. This product is mycoplasma-free and negative for many infectious disease indicators such as HIV 1&2, Hepatitis B, HCV, etc.Please refer to Figure 1 for the effect of this product in Neurobasal medium on promoting the growth of neurons in the embryonic brain of ICR mice. Figure 1. The effect of Aladdin's N-2 Supplement (100X) on neurons in the embryonic brain of ICR mice. The embryonic brain neurons isolated from ICR mice were cultured in vitro for 5 days in Neurobasal medium supplemented with this product, and immunofluorescence staining was performed with the mouse monoclonal Neuronal Class III β-Tubulin (Tuj1) Antibody which specifically recognizes neurons and the Alexa Fluor 488-labeled Goat Anti-mouse IgG (H+L) . The assay results showed that the neurons grew well and the cells interacted with each other in a high-density axonal network, partially mimicking the in vivo neuronal synapse formation to facilitate information exchange and long-term survival. This figure is for reference only, which may vary due to different experimental conditions.Please refer to Figure 2 for the effect of this product in DMEM/F12 on pro-survival of neonatal SD rat cortical neurons. Figure 2. Pro-survival effect of Aladdin's N-2 Supplement (100X) on neonatal SD rat cortical neurons. Neonatal SD rat cortical neurons isolated were cultured in vitro for 7 days in DMEM/F12 medium supplemented with this product, and then immunofluorescence staining was performed with the mouse monoclonal Neuronal Class III β-Tubulin (Tuj1) Antibody which specifically recognizes neurons and the Alexa Fluor 488-labeled Goat Anti-mouse IgG (H+L) . The results showed that neuronal survival was maintained for more than 7 days even in DMEM/F12 basal medium with relatively simplified components, and the performance of this product was comparable to that of a widely received Competitor. This figure is for reference only, which may vary due to different experimental conditions.Every 5ml of this product can be used to prepare 500ml of medium containing N-2 Supplement (1X).Precautions：This product must be used aseptically to avoid microbial contamination.This product should be protected from light. After complete dissolution and mixing well, it is recommended to be stored in aliquots in the dark at -20°C. It can also be stored temporarily in the dark at 4°C for frequent use within 1-2 weeks.A small amount of precipitation may appear after a long-term storage. It should be dissolved completely by inverting the tube and mixed well before use.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1.Add this product to the basal medium (such as Neurobasal DM DMEM/F12) at a ratio of 1:100 for the culture of neurons or other cells. Other ingredients, such as Cell Strainers (70μm, Individual Pack, Sterile) 100/box FS500 Small Animal Surgery Kit 1 kit

Specification: 100X