N2 supplement的配置和N2 Supplement操作手册

相关专题

N2培养基是细胞分化中运用的一种培养基，主要包括成份为：DMEM /F12，N2添加剂(N2 supplement )，谷氨酰胺，青霉素，链霉素， Hepes和，肝素等，N2 supplement购买的一般是gibico的，那么如何配置N2 supplement呢?N2 Supplement操作流程是什么呢?

怎样配制N2 supplement ?

偶准备养大鼠神经元细胞，以前未接触过，现在有个问题请教各位高手。买了GBICO公司的N2 supplement (5ml)，请教高手N2 supplement需要加缓冲液吗?是否直接过滤出菌就可以了?N2 supplement的包装瓶上是不是没有其所含成分的说明，谢谢。

解决方式：

GBICO的N2包装默认是无菌的,直接以培养基配至1%浓度即可.不放心的话,最好配后再过滤.包装上是没有标明配方的,需要的话我可以提供.

应该是灭过菌的，不用再过滤。一般都是100X的，建议用无菌EP管分装(分装量根据用量大小100-500微升不等)后-20保存，应用时拿出一支解冻后按所需浓度(如为100X的，则稀释100倍，即100毫升培养液里加1毫升N2)加到培养液里就行。没有说明书吗?成分说明书上肯定有。我这里给你列出：

COMPONENTS:

Human Transferrin (Holo) :转铁蛋白

Insulin (Bovine)：胰岛素

Progesterone：黄体酮

Putrescine：腐胺

Selenite：亚硒酸钠

浓度均为1mM.

注：

N-2 Supplement (100X) liquid

For the growth and expression of post-mitotic neurons and tumor Cell s of neuronal phenotype.

N-2 supplement is a chemically defined, 100X concentrate of Bottenstein's N-2 formulation (1). This supplement is recommended for the growth and expression of neuroblastomas and for the survival and expression of post-mitotic neurons in primary cultures from both the peripheral nervous system (PNS) and the central nervous system (CNS). It can be used as a substitute for the N-1 Bottenstein formulation. N-2 medium appears to be selective for neuronal Cell lines and does not support the growth of nonneuronal cell lines.

N2 Supplement作用特点以及protocol

应用

•无血清培养

•血清替代品

•神经细胞 瘤生长

•各种CNS和PNS神经元的理想配方

•与神经元基础培养基联用

Protocol for N2 Supplement

Applications

N2 Supplement is an optimised serum free supplement used in combination with B27 NeuroMix

for:

• Differentiation of ES Cell s into neuron lineage (neuron and astrocytes)

• Differentiation of neural stem cells into astrocytes and neurons

• Optimal for serum free growth for neuroblastomas

• Use in combination with B27 NeuroMix

Shelf Life and Storage

N2 Supplement is stable for 1 year when stored at ≤-15°C.

Instructions for use

N2 Supplement is a 100 fold concentrate. Dilute N2 Supplement (100x) in the base medium 1:100.

The final concentration of N2 Supplement corresponds to 1x. For preparation of 100 ml medium

add 1 ml N2 Supplement (100x) into 99 ml of the appropriate base medium.

Cell culture vessels must be coated with poly-D-lysine (0.05 mg/ml). Fibronectin must be added at

a final concentration of 5-10 μg/ml directly to the medium.

For serum free growth of neuroblastomas add N2 Supplement (100x) into Neuronal Base Medium

(supplemented with 0.5 mM L-glutamine and 25 μM glutamate) to a final concentration of 1x.

For in vitro laboratory use or further manufacturing only. Not for human use.

N2 supplement的组份在invitrogen官网上会列出，如果还需要更详细的可能只有打公司客服了，不过一般组份牵涉到产品专利。生物帮提供N2干细胞培养添加物产品 大全：N-2 Supplement ，N2 Supplement ，N-2 Plus Media Supplement ，N2 Supplement-A ，N2 Supplement-B