Ham’s F-12 Nutrient Mixture contains a wider range of components than other basal media, including zinc, putrescine, hypoxanthine and thymidine. It is often used as a basal medium for serum-free culture and is particularly suitable for single-cell culture and cloning under low serum conditions. It is also widely used for the culture of cancer cells and primary cells such as rat liver cells, rat prostate epithelial cells, chondrocytes, rat muscle cells, chicken embryo cells and more. Ham’s F-12 does not contain protein, lipids or growth factors. Therefore, it may need to be supplemented with nutrients and usually requires the addition of 10% fetal bovine serum (FBS). Ham’s F-12 uses a bicarbonate buffer system and therefore requires a 5-10% CO2 environment to maintain physiological pH value.
We offer various modified Ham’s F-12 media formulations with different components for a wide range of cell culture applications.

Ingredients of Powdered Ham’s F-12 Nutrient Mixture

P/N	Specification	Ingredient Description
HCY-BCMP09001A	1L×10 bag	Including: L-glutamine, phenol red Not contain: sodium bicarbonate
HCY-BCMP09001B	10L×1 bottle	Including: L-glutamine, phenol red Not contain: sodium bicarbonate

This Ham’s F-12 dry powder cell culture medium needs to be supplemented with sodium bicarbonate, pH adjusted and filtered during preparation. The detailed preparation plan is as follows:

1. Add 950 mL of distilled water to the mixing container as close as possible to the final volume.
2. Add the dry culture medium to room temperature (15°C to 30°C) water and stir gently. Do not heat the water.
3. Rinse the inside of the package to remove all residual powder.
4. Add 15.7mL of 75g/L NaHCO3 solution to the medium.
5. Adjust the pH value to the desired final working pH below 0.2 to 0.3 units by slowly adding and stirring 1 N NaOH or 1 N HCl. The pH value may increase by 0.1 to 0.3 units after filtration. The recommended working pH value is 7.5-7.9.
6. Adjust the final volume with distilled water.
7. Immediately use a positive pressure filtration system to filter the medium through a 0.2 μm filter membrane into a sterile container.

Ham ‘s F – 12 as the basis of serum-free culture culture, often under the low serum content, especially suitable for single cell cultivation and cloning, after adding serum is also widely used in cancer cells and the cultivation of the primary cell, such as the rat liver cells, rat prostate epithelial cells, cartilage cells, rat myoblast, chicken embryo cells, etc. In addition, the DMEM/F12 culture medium obtained by mixing Ham’s F-12 with equal volume of DMEM was more nutritious and widely used.

Ingredients of Ham’s F-12 Nutrient Mixture

Cat No.	L-Glutamine	Alanyl-Glutamine	HEPES	PSP	Penicillin-streptomycin Solution
HCY-BCM09001	★			★
HCY-BCM09001A	★			★	★
HCY-BCM09002				★
HCY-BCM09003		★		★
HCY-BCM09004	★
HCY-BCM09005	★		★	★

L-glutamine (Glutamine)

L-glutamine (Glutamine) is an essential nutrient element in cell culture, but it is unstable in solution and will degrade spontaneously. The amount of L-glutamine can be arbitrarily adjusted according to the research needs of the medium without L-glutamine, and the addition of fresh L-glutamine or its substitutes at the time of use is more beneficial to cell growth.

L-alanyl-L-glutamine (Ala-Glu), also known as alanyl-glutamine and alanyl-glutamyl dipeptide, is an advanced cell culture additive that can directly replace L-glutamine in the cell culture medium. l-Glutamine is an essential nutrient in cell culture, but it is unstable in solution and will, however, it is unstable in solution and spontaneously degrades to produce ammonia and pyroglutamic acid, of which ammonia is harmful to cells, whereas L-alanyl-L-glutamine is very stable in aqueous solution and does not degrade spontaneously. The mechanism of cellular utilization is that during cell culture, cells will gradually release a peptidase into the culture medium to hydrolyze L-alanyl-L-glutamyl into L-alanine and L-glutamine, and then cells will take up and utilize these two hydrolysis products. The process of cellular utilization of L-alanyl-L-glutamyl is similar to the flow-addition culture strategy, in that low levels of L-glutamine are continuously added to the culture medium, thereby increasing the utilization of L-glutamine without generating excess ammonia, which is more conducive to cell growth. L-alanyl-L-glutamyl can replace equimolar L-glutamine, is suitable for all cells, requires little adaptation, and can extend cell culture time. It can also extend the cell culture time and reduce the number of passages, which saves time and money. Cells cultured in medium supplemented with L-glutamine showed a slower reduction in activity than those cultured in a medium supplemented with L-glutamine. The slightly longer delay period is due to the time required for the release of peptidase and the digestion of the dipeptide.

HEPES

HEPES is an excellent biological buffer, which has no toxic effect on cells. The medium with HEPES can maintain a constant pH range for a longer period of time, which can effectively prevent cell growth from being adversely affected by large pH fluctuations of the culture medium. It can be used in CO2-free incubators.

Penicillin-streptomycin Solution

The penicillin-streptomycin solution, also commonly referred to as “dual antibiotics”, is the most commonly used antibiotic in vitro culture to prevent microbial contamination, in which penicillin interferes with bacterial cell wall synthesis and is particularly effective against Gram-positive bacteria, while streptomycin inhibits bacterial protein synthesis by binding to the 30S subunit of the bacterial ribosome and is particularly effective against Streptomycin binds to the 30S subunit of the bacterial ribosome and inhibits bacterial protein synthesis, which is effective against both Gram-negative and Gram-positive bacteria, but is particularly effective against Gram-negative bacteria. The combination of penicillin and streptomycin can prevent most bacterial contamination.

This product contains amino acids, vitamins, inorganic salts and many other components required for the multi-culture of cells, but does not contain proteins, lipids or any growth factors, so the product should be used with serum or serum-free additives.

Cautions of Ham’s F-12 Nutrient Mixture

• This product has been filtered and de-bacterized and should be used with care for the aseptic operation to avoid contamination.
• Do not freeze-thaw the product in order to maintain the best use of the product.
• This product is intended for scientific research or further study use only, not for diagnosis or treatment.