DMEM/F-12 is a 1 to 1 mixture of DMEM and Ham's F-12, combining DMEM's high nutrient concentration with F-12's broader range of components and trace elements. It supports a very wide range of cell types, often at lower serum than DMEM alone, and is the standard base for many serum-free and defined formulations. Some versions include 15 mM HEPES and some do not.
DMEM/F-12 is one of the most popular cell culture media in use today. It is a 1:1 blend of DMEM and Ham’s F-12 that combines the richness of DMEM with the wide range of components in F-12, making it a versatile choice for many cell types and for reduced-serum work. This guide explains what DMEM/F-12 is, why it is so widely used, which cells it suits, and how to use it.
Key Facts
- Type: 1:1 blend of DMEM and Ham’s F-12
- Best for: Broad cell types; reduced-serum and serum-free work
- Glucose: Intermediate (between DMEM and F-12)
- Buffering: Bicarbonate (use a CO2 incubator)
What is DMEM/F-12?
DMEM/F-12 is a basal medium made by mixing equal parts of two well-established media: DMEM, a rich, high-nutrient medium, and Ham’s F-12, which contains a broader set of components including trace elements and other additions. The blend brings together DMEM’s high concentrations of amino acids and glucose with F-12’s wider variety of ingredients, giving a balanced, well-rounded medium.
Why is DMEM/F-12 so popular?
The combination is more than the sum of its parts. DMEM supplies high levels of key nutrients, while F-12 adds a richer range of components at lower concentrations. Together they create an environment that supports a wide variety of cells and works especially well when you want to reduce or remove serum. That flexibility is why DMEM/F-12 is a default choice in so many protocols. For a direct comparison with plain DMEM, see our guide on DMEM vs DMEM/F-12.
What is DMEM/F-12 used for?
DMEM/F-12 suits a broad set of cell types, including:
- Many primary cells and established cell lines
- Reduced-serum and serum-free culture, often as a base with defined supplements
- Stem cells and specialized cultures that benefit from a rich, balanced formulation
What is in DMEM/F-12?
Because it is a 1:1 blend, DMEM/F-12 contains the salts, amino acids, vitamins, and glucose of both parent media, giving an intermediate glucose level and a broad nutrient profile that includes F-12’s trace elements. It uses a bicarbonate buffer with phenol red, so it needs a CO2 incubator (usually 5%) to hold the correct pH. HEPES-buffered versions are available for work outside the incubator.
How to supplement and use DMEM/F-12
Before use, DMEM/F-12 is completed with serum (commonly 5 to 10% FBS) or defined supplements, plus L-glutamine or a stable glutamine dipeptide. Keep it in a CO2 incubator, store it refrigerated and protected from light, warm it gently before use, and keep the same lot across an experiment for consistency. Its balanced formulation makes it a strong starting point when moving toward reduced-serum culture.
DMEM/F-12 is widely used with HEK293 cells for recombinant protein expression. For the downstream step, see our guide to His-tag protein purification.
Which DMEM/F-12 do you need?
Browse all DMEM/F-12 · all cell culture media
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Frequently Asked Questions
What is the ratio in DMEM/F-12?
DMEM/F-12 is a 1:1 blend, made by mixing equal parts DMEM and Ham's F-12. This gives an intermediate glucose level and a broad nutrient profile that includes the trace elements contributed by F-12.
Why use DMEM/F-12 instead of plain DMEM?
DMEM supplies high levels of key nutrients, while F-12 adds a wider range of components at lower concentrations, including trace elements. Together they create a richer environment that suits demanding cell types and reduced-serum or serum-free work.
Is DMEM/F-12 good for serum-free culture?
Yes. DMEM/F-12 is commonly used as a base for reduced-serum and serum-free culture, paired with defined supplements in place of some or all of the serum.
How much serum does DMEM/F-12 need?
It is typically completed with 5 to 10% fetal bovine serum, or with defined supplements for serum-free work, plus L-glutamine or a stable glutamine dipeptide. Because it is bicarbonate buffered, keep it in a CO2 incubator.
