What is the composition of fetal bovine serum?
FBS is a complex mixture of biomolecules that includes growth factors, proteins, trace elements, vitamins, and hormones that are important for the growth and maintenance of cells in vitro culture.
What are the constituents of FBS?
These components include serum, hydrolysates, growth factors, hormones, proteins, peptides, lipids, adhesion factors, and amino acids.
How to make dmem with 10 FBS?
Most cell lines I work with are cultured in standard DMEM + 10% FBS + 1% Anti/Anti. Prepare this reagent by mixing 1 bottle (500 mL) of DMEM with 50 mL of FBS and 5 mL of Anti/Anti. Shake to mix. This reagent is now ready to use and can be stored in the fridge for several weeks/months.
What is the difference between BSA and FBS?
Bovine Serum Albumin (BSA) is a purified single-type protein derived from bovine blood. Fetal Bovine Serum (FBS), on the other hand, is derived from the bovine fetus and is a complex mixture (1000+ components) of various molecules (e.g. proteins, lipids, carbohydrates, hormones, enzymes, etc.)
What is the composition of bovine serum?
BSA contains about 583 amin acid residues and does not have carbohydrates within a single polypeptide chain. It contains 17 interchain disulfide bridges and 1 sulfhydryl group at pH 5-7.
Is fetal bovine serum a protein?
FBS is rich in complex protein components, trace nutrients, and hormones required for the adhesion, growth, and proliferation of various cells (Honn et al., 1975; van der Valk et al., 2010).
What are the list of proteins in FBS?
The major component of FBS is serum albumin which constitutes 60 to 67% of total proteins. Other abundant proteins include cone cGMP-specific 3′,5′-cyclic phosphodiesterase R-subunit, alpha-1-antiproteinase, plasminogen and lactoperoxidase.
How much protein is in FBS?
Total Albumin (protein) level in FBS is less than or equal to 4.5g/dL, but this standard does not call out protein specifications for the other categories of bovine serum.
What is contained in FBS?
FBS sourced from bovine fetuses contains more than 1,000 components such as growth factors, hormones, and transport proteins that contribute to cell growth when supplemented into culture media.
Why use 10% FBS in cell culture?
We use both 10% or 5% FBS for cell culture media. Some cells require 10%, mostly for hard to culture cells or slow-growing cells (like some epithelial cells or primary cells etc.). But some cells (fast-growing cells, some cell lines etc.) like mesenchymal cells, fibroblast cells we use 5% FBS containing culture media.
What percentage is FBS for DMEM?
Therefore, DMEM requires supplementation, commonly with 10% Fetal Bovine Serum (FBS). DMEM uses a sodium bicarbonate buffer system (3.7 g/L), and therefore requires a 5–10% CO2 environment to maintain physiological pH. For Research Use or Further Manufacturing.
What is the composition of DMEM media?
Dulbecco’s Modified Eagle’s Medium (DMEM) is modified to contain 4 mM L-glutamine, 4500 mg/L glucose, 1 mM sodium pyruvate, and 1500 mg/L sodium bicarbonate. It has applications for cell culture, cell growth, and viability.
How to make 10% BSA?
If one wants to make a 10% (100mg/mL) stock solution of BSA powder, they must dissolve 1 g BSA powder in 10 mL of distilled water. It is also important to avoid clumping, by layering the powder on the surface of the liquid when dissolving BSA powder.
Can BSA replace FBS?
Yes, the 1% BSA may be substituted with 10% FBS for plate blocking. Dilution of the standard, serum samples and detection antibody, in buffers containing 10% FBS may be preferable for testing of serum samples or tissue culture fluids.
What is BSA protein standard?
BSA is the universally accepted reference protein for total protein quantitation. The albumin standard is precisely formulated at 2 mg/mL in an ultrapure 0.9% sodium chloride (saline) solution.
How to make fetal bovine serum?
Cows pass both veterinary pre- and post-mortem inspections. During extraction, blood is drawn via cardiac puncture from the expired fetus in a closed system using best practices to regulate hemoglobin and endotoxin levels. Blood is refrigerated to encourage clotting and processed into raw serum.
Why is BSA used in protein estimation?
Using BSA as a Protein Standard It is also frequently used as a reference standard for protein assays, as BSA is readily available, easy to purify, highly stable, and lacks enzymatic activity.
What is the concentration of protein in FBS?
The overall amount of protein levels in FBS is 50% that of human plasma protein, and since cell culture media contains 5%–10% FBS, the protein binding of nicotine, ceftizoxime, and moxifloxacin can be considered negligible for in vitro testing (Figure 1).
What is the lipid composition of fetal bovine serum?
FBS has one-quarter the lipid content, one-third the MUFA, 11% of the PUFA and 8% of the normal level of linoleic acid [9, 10] compared to media made from human serum (see Table 1 for the relative fatty acid composition of sera and plasma of different vertebrates).
Does fetal bovine serum contain albumin?
The globular protein, bovine serum albumin (BSA), is a major component of fetal bovine serum. The rich variety of proteins in fetal bovine serum maintains cultured cells in a medium in which they can survive, grow, and divide.
Does FBS contain amino acids?
Whereas standard FBS will contain higher concentrations of small molecules (e.g., amino acids, hormones, cytokines), the careful filtration process used to produce dialyzed FBS greatly reduces the concentrations of these molecules, facilitating more precise control of nutrients and cell signaling molecules in the cell …
What molecules are in FBS?
FBS is a mixture formed by removing fibrin from fetal bovine plasma, which contains various plasma proteins, peptides, fats, growth factors, hormones, inorganic substances and some unclear small molecules, including different kinds and concentrations of metabolites that may have great effects on the biological …
Is FBS a carrier protein?
The most commonly used carrier proteins include bovine serum albumin (BSA), human serum albumin (HSA), or fetal bovine serum (FBS). These carrier proteins are generally used at 0.1% concentration.
What is the difference between bovine calf serum and FBS?
The newborn calf serum is generally more stable in price and availability compared to FBS if reliability is a key concern. Newborn calf serum is useful in applications where the cell line is robust and cell attachment is the primary goal.
Does FBS contain IgG?
Ultra-low IgG fetal bovine serum has been chromatographically depleted of IgG, an important factor in isolating organisms that may be inhibited by pre-existing levels of IgG or for quantitative IgG immunoassays.
How is FBS made?
Cows pass both veterinary pre- and post-mortem inspections. During extraction, blood is drawn via cardiac puncture from the expired fetus in a closed system using best practices to regulate hemoglobin and endotoxin levels. Blood is refrigerated to encourage clotting and processed into raw serum.
What is the composition of horse serum?
Donor Horse Serum contains more immunoglobulins than fetal bovine serum (FBS) and has an increased protein content. It is a cost-effective alternative to fetal bovine serum. Donor Horse Serum involves a controlled collection of the raw serum, as closed herds can be used.
What is serum derived bovine immunoglobulin made of?
Serum derived bovine immunoglobulin/protein isolate (SBI) is approximately 92% protein (of which >50% is IgG), and is free of milk, soy, and gluten products [28, 63].
Does FBS contain glucose?
Thermo fisher FBS contains less than 50 mg/L of glucose and using the same should not affect your result.
What is fetal bovine serum?
Can fetal bovine serum be used for cell culture?
What is fetal bovine serum (FBS)?
What is a globular protein in fetal bovine serum?
Fetal bovine serum (FBS) is a vital component in many cell culture labs, providing essential nutrients, growth factors, and hormones to keep cells happy and growing. But what exactly is in FBS? What makes it so special? Let’s dive into the world of FBS protein composition!
A Diverse Protein Mix: The Key to Cell Culture Success
FBS is a complex mixture of proteins, with over 1,500 different proteins identified so far. That’s a lot of variety! This diverse protein cocktail is what makes FBS so valuable in cell culture. These proteins play crucial roles in:
Cell Growth and Proliferation: FBS contains a whole host of growth factors, like insulin-like growth factor 1 (IGF-1), platelet-derived growth factor (PDGF), and epidermal growth factor (EGF), which stimulate cell division and proliferation.
Cell Survival and Protection:Antioxidant proteins and cytokines in FBS help protect cells from damaging free radicals and inflammation.
Cell Attachment and Adhesion:Extracellular matrix proteins, like fibronectin and laminin, help cells stick to the culture dish and form healthy cell layers.
Hormonal Support: FBS provides essential hormones, like thyroid hormone and steroid hormones, to regulate cell function and metabolism.
Breaking Down the Protein Lineup: Major Players in FBS
While FBS is a complex mix, some key protein families stand out. Let’s take a closer look:
Albumin: This protein is the most abundant in FBS, acting as a carrier protein for various molecules, including hormones and fatty acids.
Immunoglobulins: These antibodies help protect cells from infection and contribute to the immune system of the serum.
Transferrin: This iron-binding protein ensures cells have access to this essential mineral for growth and development.
Growth Factors: We already mentioned these key players, but it’s worth emphasizing their importance again. Growth factors are critical for promoting cell division and proliferation, making them vital for healthy cell cultures.
Understanding the Variability: Why FBS Batches Can Differ
FBS is a natural product, and its protein composition can vary depending on several factors, including:
Source of the serum: The age of the fetus, the breed of cattle, and even the season of collection can influence the protein profile of FBS.
Processing methods: Different manufacturers employ distinct techniques to extract, purify, and sterilize FBS, which can lead to variations in protein content.
This variability is why it’s crucial to choose a high-quality FBS from a reputable supplier and be mindful of lot-to-lot variations. Always test your cells with different FBS batches to ensure consistent results.
Going Beyond the Basics: Advanced Techniques for Understanding FBS Protein Composition
Scientists are constantly refining their understanding of FBS protein composition. Advanced techniques, like proteomics, are allowing us to:
Identify and quantify even more proteins present in FBS.
Understand how protein expression changes under different conditions, such as during cell culture.
Develop better FBS-free alternatives, such as protein-defined media, that provide the essential nutrients and growth factors for specific cell types.
FBS: A Complex and Essential Tool for Cell Culture
Fetal bovine serum is a vital tool in cell culture, offering a complex mix of proteins that support cell growth, survival, and function. Understanding the composition and variability of FBS is essential for obtaining consistent and reliable results in your cell culture experiments.
As research into FBS protein composition continues, we can expect even more insights into its benefits and potential limitations, paving the way for more refined cell culture techniques and improved therapeutic applications.
FAQs:
Q: What is the difference between FBS and FCS?
A: FBS and FCS both refer to fetal bovine serum. The abbreviation can vary depending on the region or the supplier.
Q: Why is FBS so widely used in cell culture?
A: FBS provides a rich source of essential nutrients, growth factors, hormones, and proteins that support cell growth and survival. It’s a cost-effective and well-established solution for many cell culture applications.
Q: Is there a way to reduce or eliminate the use of FBS?
A: Yes, there are efforts to develop FBS-free media, using defined protein mixtures or alternative growth factors. These alternatives offer more consistency and address ethical concerns related to using animal-derived products.
Q: Are there ethical concerns related to using FBS?
A: Yes, the collection of FBS involves the harvesting of fetal calves, raising ethical concerns about animal welfare. This is one of the driving forces behind the development of FBS-free alternatives.
Q: How can I find a high-quality FBS for my cell culture experiments?
A: It’s important to choose a reputable supplier who offers FBS that has been thoroughly tested for sterility, endotoxin levels, and other quality parameters. Look for suppliers that provide detailed information about the origin of their FBS and the processing methods used.
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