B-27™ Supplement (50X), serum free, 10 mL - FAQs

做为一般性的参考方案，我们推荐您使用Gibco B-27添加剂来培养神经干细胞，海马及其他类型的CNS神经元。

-对于来自PNS和CNS中的神经母细胞瘤或有丝分裂后的神经元，可添加Gibco N-2添加剂。
-对于原代胶质细胞（星形胶质细胞）或具有星形胶质细胞表型的肿瘤细胞系（星形胶质细胞和胶质细胞瘤），或少突胶质细胞，可使用Gibco G-5添加剂。

Yes, the protein content in this product is high enough so filtering through a low protein binding filter as a 50X or 1X in solution should not be a problem.

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No, it is not recommended to freeze media due to the potential of precipitates forming upon thaw. Inorganic salts and amino acids in the formulation may come out of solution when exposed to temperature fluctuations. These precipitates will not go back into solution easily once formed.

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Neurobasal and Neurobasal-A media (for postnatal and adult neurons) allow for long-term maintenance of neuronal cells without the need for an astrocyte feeder layer. These media should be supplemented with either serum or a serum-free supplement, plus 0.5mM L-glutamine. B-27 supplement is a serum-free supplement that comes as a 50X concentrate in a 10ml volume. This is enough supplement for 500ml of media. Fetal, postnatal, and adult neural cultures can be grown in the appropriate Neurobasal medium supplemented with B-27 supplement .

We also have two other supplements. One is called G-5 and is for growth and expression of glial cells (normal and tumor) of astrocytic phenotype. This comes in a 1ml size, at a 100X concentration. The other supplement is called N-2 and is for growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype. This comes in a 5ml size, at a 100X concentration.

For more information on Neurobasal media, search "Neurobasal" from our website home page.

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We recommend using Neurobasal Plus Medium with B-27 Plus Supplement for long-term culture of mixed hippocampal cells. For short-term culture of a pure population of hippocampal neurons, we would recommend using Neurobasal-A Medium with B-27 Supplement.

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Yes, we do offer B-27 Supplement, minus insulin (Cat. Nos. A1895601 and A1895602). All our other B-27 supplements do contain insulin.

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Once B-27 Supplement has been added to a basal medium, the medium should be used within 2 to 4 weeks. If it will take longer than that to use up the medium, we recommend that you aliquot the B-27 Supplement and supplement smaller volumes of medium as needed.

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B-27 Plus Supplement is an optimized formulation of classic B-27 Supplement with a streamlined manufacturing process. No small molecules or growth factors have been added.

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We do not recommend thawing at 37 degrees C. Best results are observed upon thawing at room temperature for approximately 2 hrs or overnight at 2-8 degrees C.

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We recommend thawing it at 4 degrees C overnight or at room temperature for 2 hours. We don't recommend thawing in a water bath or incubator.

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We recommend using B-27 Plus Supplement and Neurobasal Plus Medium instead of the regular versions of B-27 Supplement and Neurobasal Medium products (not specialty versions like minus Insulin, minus Vitamin-A or Neurobasal-A) in protocols used for:

- Maintenance of primary rat, mouse and human PSC-derived and fetal-derived neurons
- Differentiation of human PSC-derived and fetal-derived neural stem cells (NSCs) to neurons

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Yes, we recommend that you aliquot when you first thaw it and thereafter subject the aliquot to no more than 2 additional freeze/thaw cycles.

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We offer the following B-27 supplement products, which are developed for specific applications:

- B-27 Serum-free Supplement (Cat. No. 17504044): Complete formulation for the growth of long-term viability of neurons.
- B-27 Supplement minus AO (antioxidants) (Cat. No. 10889038): Five antioxidants are removed. The supplement is ideal in studies of oxidative damage, apoptosis, or applications in which free radical damage to neurons occurs.
- B-27 Supplement minus Insulin (Cat. No. A1895601): Insulin is removed. The supplement is ideal for studies of insulin secretion and insulin receptors.
- B-27 Supplement minus Vitamin A (Cat. No. 12587010): Vitamin A which induces neural differentiation is removed. The supplement is ideal for proliferation of stem cells.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

As a general guideline, we recommend using Gibco B-27 Plus Supplement to culture neural stem cells, hippocampal, and other CNS neurons.

- For neuroblastomas, or post-mitotic neurons from both PNS and CNS, Gibco N-2 Supplement can be used.
- For primary glial cells (astrocytes) or tumor cell lines of astrocytic phenotype (astrocytes and gliomas), or oligodendrocytes, Gibco G-5 Supplement can be used.