Fetal Bovine Serum, Premium Plus, bottle, 500 mL - FAQs

我们的研究结果显示在4°C条件下短时保存化冻的FBS，28天（之内）不会降低其生长或活性方面的效能。如需在4°C条件下将FBS保存更长时间，就需要逐案评估这一储存条件是否为必须。

多种原因可能导致FBS中出现絮状沉淀。最为常见的原因是血清中脂蛋白的变性。你可能会在血清化冻后观察到血纤蛋白——一种血清中存在的促凝蛋白。这并不会影响本品的使用性能。

如需去除絮状沉淀，请将血清移至无菌管中，以400 x g进行简短离心。之后把所得的上清液和培养基一并过滤。不要试图过滤含有絮状沉淀的血清，可能会堵塞滤器。

Gibco FBS未经预老化处理。当储存于2至8°C的条件下，血清中可能存在的各类蛋白与脂蛋白（如冷凝集素，纤维蛋白原，玻连蛋白等）会发生聚集，并且形成肉眼可见的沉淀或混浊。这并不会影响血清的使用性能。我们推荐您将FBS储存于–20°C，同时避免反复冻融。

通常确认是否需要此操作的最好办法是咨询您获取细胞的来源。举例来说，ATCC对于他们所销售的细胞就有血清方面的要求。多数昆虫细胞系与胚胎干细胞系都需要应用热灭活的血清。

该蛋白是一个能够反应快速收集和加工效果的标记物。

这些FBS在进行透析时通过使用分子量为10000的分子筛在0.15M NaCl溶液中进行正切流动过滤，直至葡萄糖水平低于5 mg/dL。由于这一操作非彻底透析，因此氨基酸一类的低分子量透析组份可能无法完全去除。这些产品未经彻底透析的原因是该操作可能会导致血清中肽类的沉淀和失活。

BVD会造成牛的病毒性腹泻。该病症是牛最为常见的病毒感染症状之一。据估计在世界范围内，牛种群中估计有百分之七十至九十的个体血清为BVD阳性。该病毒会导致牛发病和流产。也有部分现存的BVD毒株是非致病性的。
在细胞培养应用中，联邦法规（9CFR）对生物制品生产所用细胞系和原料提出了一些要求。用于生产生物制品的血清要求具有阴性的BVD病毒检测结果。
Gibco血清的BVD病毒测试均按照9CFR中的检测方案来进行。我司内部划分为0至4级（根据检测结果）。结果为0意味着遵照9CFR测试方案，本批血清的病毒检测结果为阴性。1-4级的检测结果表明本批血清为病毒阳性。A+1表明病毒发出较低荧光，a+4表示病毒荧光较强。该批次的质检报告中会标记“经测试（tested）”一词，而非实际的BVDV抗体荧光强度。
某一批次的血清产品按照9CFR方案测试结果为0，而在使用PCR一类更新更灵敏的技术测试时则可能出现阳性结果。为了确保该批次血清“真正”为BVD病毒阴性，需对其进行伽马辐照等的处理。

Gibco FBS有100 mL、500 mL和1000 mL的瓶装规格，也有3.5 L和4 L体积的产品可供特殊订购。

您可将解冻后的产品置于56°C的恒温控制下孵育30分钟。我们所提供塑料和玻璃瓶装的血清产品均能胜任热灭活处理。警告：不要试图以更高的温度和更长的时间来热灭活血清产品，这样的条件会导致蛋白变性，进而降低产品性能。

热灭活步骤能够灭活补体系统。补体会参与以下事件中：溶解细胞事件，平滑肌收缩，肥大细胞与血小板释放组胺，吞噬作用增强，趋化作用，淋巴细胞与巨噬细胞的活化等。

监管机构通常将伽马辐照作为灭活动物血清中天然存在的病毒，以及降低病毒风险的最佳方法。我们以现货提供少量的伽马辐照产品。我们大多数的标准目录产品均可依照客户的要求来定制伽马辐照操作。我们所提供的定制工艺覆盖了多种照射剂量范围，并经过完整的效果验证。

伽马辐照被认为是灭活动物源性产品中病毒的有效手段。按照USDA（美国农业部）对抗体产品常规要求方面的规定（9CFR，章节113.450），动物源性血液衍生物的最小照射剂量为25 kGy。某些欧洲国家要求在进口前对产品进行处理的最小剂量为25 kGy。
我们将依照您的需求对血清产品进行伽马辐照。我们通过经验证的30-45 kGy的伽马辐照来灭活可能存在于FBS中的最为常见的牛病毒和支原体。病毒灭活的对数（log）量级达6-8，支原体灭活的对数量级达6-7。我们也验证了血清的生化性质与细胞培养性能不会受30-45 kGy伽马辐照的影响。

有的。我们为您提供了各种的定制选择。请联系我们的血清专员来讨论您的应用与研究需求。我们将通过密切合作来为您提供专项加工和测试的血清产品，以匹配您的特定需求。举例来说，我们的定制包括热失活、伽马辐照和四环素残留测试等。

不必，使用前并不需要过滤血清。Gibco血清产品经最为严格的工艺生产，其中就包含有膜过滤步骤。

Fetal Bovine Serum（FBS）一度被称作Fetal Calf Serum（FCS）。它们是同种产品。

血清浓度随所用细胞系和基础培养基的不同而变化。请点击此处(https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/advanced-d-mem-and-mem/recommended-sera-supplementations.html)查看我们推荐用于测试细胞系的血清添加剂。

会。动物血清应储存于–5至–20°C。培养基储存于2至8°C；请在推荐的保质期内用完。请将完全培养基（加入添加剂）保存于2 至 8°C，推荐的存放期限为2至4周。此外，尽量减少血清和培养基暴露于光线下的时间。

1. Heating inactivates complement. Active complement can participate in cytolytic events, contract smooth muscle, release histamine from mast cells and platelets, and activate lymphocytic and macrophage cells. Applications where heat-inactivated serum is recommended include immunological studies and culturing of embryonic stem cells (ESCs), insect cells, and smooth muscle cells.
2. Heat inactivation helps to achieve bottle-to-bottle and lot-to-lot stability by neutralizing many factors that can vary largely from lot to lot.
3. There aren't necessarily disadvantages to heat inactivation of FBS, but there is some evidence that suggests there may be no added benefit to it unless you are carrying out immune studies.

Note: Heat inactivation is performed in a 56 degrees C water bath for 30 min with swirling every 10 min or so for heat distribution and to lower the degree of protein aggregation/flocculant precipitation. Note: If the time or temperature is exceeded, the serum may thicken to a gel. If this occurs, the serum is no longer usable. Unnecessary heat inactivation can take up time and potentially lead to wasted reagents if a mistake is made during the protocol¹.

1. Pellerin, et al., Bioengineering, published in 2021.

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Our CoAs only capture the total protein content not specifically the amount attributed to albumin.

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FBS is added to culture medium at a concentration of 2-10% to provide attachment factors, nutrients, and hormones for mammalian cells, as well as to be a buffer against disruptions like pH changes and endotoxins. FBS has significant amounts of embryonic growth promoting factors like hormones, carrier proteins, and macromolecular proteins. It also has low levels of antibodies and other growth-inhibiting components. For most basal medium, 10% FBS is used. Some applications require up to 20% supplementation, so researchers need to determine what is optimal for their specific application and cell line.

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This product can be stored at 4 degrees C for up to 4 weeks. Once the medium has been supplemented with serum, we recommend using it within 2-4 weeks.

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We recommend thawing the serum overnight at 4 degrees C or in a 37 degrees C water bath, removing as soon as it is thawed. Once thawed, aliquot into single-use sizes and freeze the aliquots. Each aliquot should ideally be thawed only one additional time as repeated freeze-thaw cycles are not recommended.

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The product is received frozen and should be stored at -5 degrees C to -20 degrees C right away. Do not subject the serum to repeated freeze-thaw cycles.

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Yes, we do offer Fetal Bovine Serum, certified, heat-inactivated ).

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Heating inactivates complement. Active complement can participate in cytolytic events, contract smooth muscle, release histamine from mast cells and platelets, and activate lymphocytic and macrophage cells. Applications where heat-inactivated serum is recommended include immunological studies and culturing of embryonic stem cells (ESCs), insect cells, and smooth muscle cells.

Heat inactivation is performed in a 56 degrees C water bath for 30 min with swirling every 10 min or so for heat distribution and to lower the degree of protein aggregation/flocculant precipitation. Note: If the time or temperature is exceeded, the serum may thicken to a gel. If this occurs, the serum is no longer usable.

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Fetal bovine serum is triple filter sterilized using a 0.1 µm filter. All our Gibco cell culture liquid products are prepared by an aseptic process for which each step has been validated to ensure that all products meet the industry standard sterility assurance level of 10^-3; i.e., product that demonstrates a contamination level of no more than 1 of 1,000 units during the manufacturing process. The highest level of sterility assurance (equal to or greater than 10^-6) cannot be achieved without terminal sterilization which is harmful to the performance of cell culture products. Key points of control include validated sterilization cycles for all material with product contact, routine media fills employing bacteriological media, a comprehensive environmental monitoring program, validated cleaning procedures, and a validated final filter integrity testing program. Additionally, filtration and dispensing are performed within positive pressure, HEPA-filtered, environmentally controlled rooms.

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Since our sera are not pre-aged before filtration, turbidity or flocculent debris may develop during thawing or storage. This flocculence is a normal characteristic of the serum and does not adversely affect performance characteristics of the serum.

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No, the tetracycline testing is not being performed for serum lots manufactured after March 1, 2019. This testing can be requested as a custom request.

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You can test each new lot to be sure that it works for your specific applications. Alternatively, our serum sales team can determine the most consistent, highest-performing serum lot available using the Gibco iMatch Sera Lot Matching Tool.

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Yes, after the overnight thaw, you can aliquot the serum into smaller, single-use aliquots and refreeze it.

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Fetal bovine serum is triple-filtered through 0.1 µm filters. Bovine serum is filtered through 0.2 µm filters.

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Heating serum inactivates complement. Active complement can participate in cytolytic events, contract smooth muscle, release histamine from mast cells and platelets, and activate lymphocytic and macrophage cells. Applications where heat-inactivated serum is recommended include immunological studies and culturing of embryonic stem cells (ESCs), insect cells, and smooth muscle cells.

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Serum can be kept at 2-8 degrees C for up to 4 weeks.

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Our studies have shown that short-term storage of thawed FBS at 4 degrees C for up to 28 days causes no decrease in growth or viability performance. FBS stored at 4 degrees C for longer periods of time should be evaluated on a case-by-case basis should this storage condition be necessary.

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Flocculence may appear in FBS for a variety of reasons. The most common reason is the denaturation of serum lipoproteins. You may observe fibrin, one of the clot-forming proteins present in serum, after the serum has been thawed. This should not affect product performance. To remove the flocculence, transfer the serum to sterile tubes and centrifuge the material briefly at 400 x g. Then filter the resulting supernant along with your media. Do not attempt to filter serum containing flocculence, it may clog filters.

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Gibco FBS is not pre-aged. When stored at 2 to 8 degrees C, the possibility exists for various proteins and lipoproteins in serum (e.g., cold agglutinins, fibrinogen, vitronectin, etc.) to aggregate, and form either perceptible material or observed turbidity. This should not affect serum performance. We recommend that you store FBS at -20 degrees C and avoid repeated freeze-thaw cycles.

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Usually the best way to find out is to go to the source you obtained your cells from. For example, ATCC will have serum requirements for the cells they sell. Most insect cell lines and embryonic stem cell lines require heat-inactivated serum.

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It is a quality indicator that reflects rapid collection and processing.

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It is dialyzed by tangential flow filtration against 0.15 M NaCl using a 10,000 molecular weight cut-off membrane until the glucose level is less than 5 mg/dL. Since this is not exhaustive dialysis, low-molecular weight dialyzable components, such as amino acids may not be totally removed. Exhaustive dialysis is not performed because it can result in precipitation and inactivation of serum peptides.

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BVD stands for Bovine Viral Diarrhea. It is one of the most common viral infections in cattle. It is estimated that 70 to 90 percent of the world's cattle population is seropositive for BVD. This virus can cause abnormalities and fetal abortions in cattle. Several strains of BVD exist, some of which are non-pathogenic.

Bovine serum is tested in accordance with 9 CFR, Section 113.53. The BVDV fluorescent antibody test is one of the required tests to meet Title 9 part 113.53 of the Code of Federal Regulations. However, the results of this test are somewhat subjective in the way they are scored. They are scored 0 to +4, based on the level of observed fluorescence. Most samples will have a detectable level of BVDV antigen due to its prevalence in the bovine population. The FA part of the testing can reveal the presence of non-cytopathic BVD strains. The Cytopathogenic and Hemadsorbing Agents testing is used to determine the release of bovine serum, regardless of the BVDV result. We currently report the BVDV results as “Tested”. Any live cytopathic bovine viruses (including the cytopathic BVDV strain) would be revealed in the testing for cytopathogenic agents. In this portion of the testing, the cultures are microscopically monitored for evidence of inclusion bodies, abnormal number of giant cells, or other cytopathology indicative of cell abnormalities. The hemadsorption assay detects the presence of hemagglutinating viruses. The viral hemagglutinin would induce clumping of red blood cells. If a positive result in the cytopathogenic agents or hemadsorbing agents assays is reported, the material would be failed and therefore would not be released by Quality Assurance.

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Gibco FBS is available in 50 ml, 500 mL, and 1,000 mL bottles and by special order in 3.5 L and 4 L volumes.

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You can incubate the thawed product at a thermostatically controlled temperature of 56 degrees C for 30 minutes. Serum can be heat-inactivated in both our plastic and glass bottles. Warning: Do not attempt to heat-inactivate at a higher temperature for prolonged periods as this may compromise the product's performance through protein denaturation.

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The heating process inactivates portions of the complement cascade. Complement occurs in the following events: cytolytic activities, contraction of smooth muscle, release of histamine from mast cells and platelets, enhanced phagocytosis, chemotaxis, and activation of lymphocytic and macrophage cell types.

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Gamma irradiation is generally recognized by regulatory agencies as the best method of virus inactivation and for reducing the risk of viruses that are naturally present in animal sera. We offer a small range of gamma-irradiated products ex-stock. Gamma irradiation is also available on request as a custom service for most standard catalog products. We offer a range of fully validated custom processes covering many dose ranges.

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Gamma irradiation is recognized as an effective method for inactivating viruses in animal-origin material. Based on USDA regulations for the general requirements for antibody products (9CFR, Section 113.450), the minimum dosage for blood derivatives of animal origin is 25 kGy. Certain European countries require products to be treated prior to importation with a minimum dose of 25 kGy. We will gamma-irradiate serum on request. We have validated a process for utilizing gamma irradiation in the range of 30-45 kGy to inactivate the most common bovine viruses and mycoplasmas that may be present in FBS. The level of inactivation is 6-8 logs for viruses and 6-7 logs for mycoplasmas. We have also demonstrated that physiochemical properties and cell culture performance of serum is not altered by gamma irradiation at levels of 30-45 kGy.

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Yes. We offer a wide range of custom options. Please contact techsupport@thermofisher.com to find your Sera Specialists to discuss your application and research needs. We will work with you to supply sera that is specially processed and tested to meet your individual requirements. Some examples of our custom capabilities are heat inactivation, gamma irradiation, and testing for tetracycline residues.

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No, there is no need to filter the serum before use. Gibco serum products are manufactured using the most stringent processes, including membrane filtration.

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There is no difference in the general use of the terms. Both terms refer to serum from an unborn cow. Fetal Bovine Serum (FBS) was once known as Fetal Calf Serum (FCS).

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The serum concentration will vary with the cell line and basal medium used. Please go here to see our recommended sera supplementations for tested cell lines (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/advanced-d-mem-and-mem/recommended-sera-supplementations.html).

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Yes. Store animal sera at -5 to -20 degrees C. Store media at 2 to 8 degrees C; use within recommended shelf life period. Store complete media (supplemented) at 2 to 8 degrees C, and for complete medium the recommended shelf life is 2 to 4 weeks. Additionally, minimize exposure of sera and media to light.