Products for Media Formulation
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Cell
culture is essential to biotechnology. Historically, cells have been
cultured in serum containing media. Serum is a source of many nutrients
required by cells to proliferate and survive. However, the use of serum is
problematic. It is complex, expensive and a potential source of adventitious
biological contaminants.
Eliminating
the use of serum in cell culture requires an understanding of what it does in in
vitro systems. Among other things, serum contains proteins which serve as
carriers and protective agents for other molecules. Albumin is a protein
in serum which serves these and other functions. Albumin binds vitamins
such as pyridoxal, fatty acids, such as oleic, linoleic, linolenic, arachidonic,
myristic and palmitic acids, and ions such as copper. It increases the
solubility of these molecules, maintains them in chemical states that are
unstable in an unbound form, and protects them from oxidation.
Sigma
offers a number of albumins for use in formulating cell culture media.
Some albumins are purified using methods that protect the natural association
of albumin and the bound molecules. Others are stripped of these molecules so
that the may be reconstituted in a defined way.
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| Product # |
Product Name |
Description |
Application |
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A 1470 |
Albumin Cohn
AnalogTM |
- Powder;
- Purity, minimum 95%;
- Low endotoxin;
- Cell culture tested.
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Recommended as a growth-enhancing supplement
for cell culture. Manufactured by a novel process that maintains the lipid and growth factor characteristics of cold-ethanol precipitated
albumins.
Fatty acids which are naturally associated with albumins are important for the growth
of many biotechnologically useful cells lines. Albumin serves as natural delivery molecule for free fatty acids such as oleic,
linoleic, linolenic, and arachidonic acid.
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A 1595 |
Albumin Solution (10%) Fraction V
Bovine |
- Liquid;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Recommended as a base for the preparation
of a defined, fatty acid supplemented albumin. Can be loaded with up to 4 moles of fatty acids per mole of albumin.
Fatty acids are important for the growth of many biotechnologically
useful cells lines. Albumin serves as natural delivery molecule for free fatty acids such as oleic, linoleic, linolenic, and arachidonic acid.
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| A 9576 |
Albumin Solution (30%) Fraction
V
Bovine |
- Liquid;
- Prepared from A 9418 in D-PBS (D 5652);
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Recommended as a base for the preparation
of a defined, fatty acid supplemented albumin. Can be loaded with up to 4 moles of fatty acids per mole of albumin.
Fatty acids are important for the growth of many biotechnologically
useful cells lines. Albumin serves as natural delivery molecule for free fatty acids such as oleic, linoleic, linolenic, and arachidonic acid.
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I 3153 |
Iron Supplement, Chemically Defined (1000x)
Animal Component-free. |
- Liquid;
- Proprietary product;
- Recommended for use in cell culture applications at 0.5 to 2 ml per liter of
medium.
- Iron content: 222-334 ppm
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Designed
to replace transferrin for iron transport and delivery to cells in a broad
range of cell culture applications.
The proper
sequestration of iron in media is a critical factor in preventing oxidative
stress to cells. Iron is an essential grow element for all cell systems.
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Many
cells require hormones to modulate their proliferation or differentiation.
Sigma
offers a wide range of hormones formulated and tested for use in cell
culture. This table provides a partial list of hormones or hormone
formulations that may be particularly useful in biotechnology applications.
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| Product # |
Product Name |
Description |
Application |
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H 6909 |
Hydrocortisone Solution Animal Component-free |
- Liquid;
- Formulated to supplement at 0.2 to 2 ml per liter of medium;
- Formulated to contain 50 ug/ml in cell culture grade water (W 3500);
- Sterile filtered
- Endotoxin tested;
- Cell culture tested.
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Use as a cell culture medium
supplement.
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I 2767 |
Insulin, Human Recombinant,
sodium salt Escherichia coli |
- Powder;
- FW: 5749.5;
- Purity, not less than 90% by HPLC;
- Soluble in 10 mM HCl (20 mg/ml);
- Zinc, not more than 0.01%.
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A two-chain polypeptide hormone
produced by the beta-cells of pancreatic islets. Its molecular weight is approximately 5800 Da. The alpha and beta chains are
joined by two inter-chain disulfide bonds, the alpha chain contains an intra-chain disulfide bond. Insulin regulates the
cellular uptake, utilization, and storage of glucose, amino acids and fatty acids and inhibits the breakdown of glycogen, protein
and fat.
Recombinant insulin in the sodium salt form for use in medium supplementation. Insulin is widely used in media formulations. Suitable
for use in serum-free media formulations when the use of animal-derived proteins
is to be avoided.
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I 0259 |
Insulin, Human Recombinant, Escherichia
coli |
- Powder;
- FW: 5807.6;
- Potency: not less than 27.5 USP units/mg on a dry basis;
- Soluble in 10 mM HCl (20 mg/ml).
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A two-chain polypeptide hormone
produced by the beta-cells of pancreatic islets. Its molecular weight is approximately 5800 Da. The alpha and beta chains are
joined by two inter-chain disulfide bonds, the alpha chain contains an intra-chain disulfide bond. Insulin regulates the
cellular uptake, utilization, and storage of glucose, amino acids and fatty acids and inhibits the breakdown of glycogen, protein and fat.
Insulin is widely used in media formulations. Suitable for use in serum-free media formulations when the use of animal-derived proteins is to be avoided.
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I 0516 |
Insulin
Solution, Bovine derived, natural |
- Liquid;
- Formulated to supplement at 0.2 to 2 ml per liter of medium;
- Formulated to contain 50 mg/ml in 25 mM HEPES, pH 8.2;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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A two-chain polypeptide hormone
produced by the beta-cells of pancreatic islets. Its molecular weight is approximately 5800 Da. The alpha and beta chains are
joined by two inter-chain disulfide bonds, the alpha chain contains an intra-chain disulfide bond. Insulin regulates the
cellular uptake, utilization, and storage of glucose, amino acids and fatty acids and inhibits the breakdown of glycogen, protein
and fat.
Insulin is widely used in
media formulations. Ready to use solution
of insulin. Suitable for use in serum-free media formulations when animal-derived components are acceptable.
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I 9278 |
Insulin
Solution, Human Recombinant Escherichia
coli |
- Liquid;
- Formulated to supplement at 0.5 to 1 ml per liter of medium;
- Formulated to contain 10 mg/ml in 25 mM HEPES, pH 8.2;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested, CHO, L929, and BHK.
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A two-chain polypeptide hormone
produced by the beta-cells of pancreatic islets. Its molecular weight is approximately 5800 Da. The alpha and beta chains are
joined by two inter-chain disulfide bonds, the alpha chain contains an intra-chain disulfide bond. Insulin regulates the
cellular uptake, utilization, and storage of glucose, amino acids and fatty acids and inhibits the breakdown of glycogen, protein
and fat.
Insulin is widely used in media formulations. Ready to use solution of insulin. Suitable for use in media formulations when the
use of animal-derived proteins is to be avoided.
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Traditionally,
media formulations are based upon the addition of individual L-amino
acids. These are often referred to as essential and non-essential amino
acids. It is now understood that cells can also use amino acids that are
in the peptide form. Peptides can range in size from two amino acids to many
amino acids. The size and make-up of the peptide mixture is believed to affect
their availability to cells. Peptides that are derived by the hydrolysis of proteins provide a supplemental
(undefined) source of amino
acids in a form that supports superior growth of a number of cell types.
Sigma
offers hydrolysates from various non-mammalian protein sources to provide different mixtures
of amino acids in peptide form for use in cell culture.
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| Product # |
Product Name |
Description |
Application |
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H 6784 |
HyPep
4601TM Protein Hydrolysate
Wheat gluten derived Animal component-free. |
- Powder;
- Formulated to supplement at 1 to 5 gm per liter of medium;
- Endotoxin tested, by supplier;
- Cell culture tested.
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Recommended for use as a rich source of L-glutamine and other small peptides in
cell culture systems when the use of animal-derived proteins is to be avoided.
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P 8358 |
Peptone, Fish Derived. |
- Powder;
- Formulated to supplement at 1 to 5 gm per liter of medium;
- Cell culture tested.
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This supplement is useful for delivering additional amino acids, in the form of
small peptides, to the cell culture medium. Supplementation of a medium with
peptides often results in superior cell proliferation.
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P 0521 |
Peptone, Type IV;
Soybean derived. (HySoyTM) |
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This is an enzymatic hydrolysate of soybean
protein useful in microbial fermentation and mammalian cell culture. Supplementation of a medium with
peptides often results in superior cell proliferation.
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Lipids
are essential to the proliferation, growth and survival of all cells.
Among the critical building blocks of lipids are cholesterol and the fatty
acids.
Cholesterol
is a major component of all biological membranes (approximately 25% of the total brain
is cholesterol), and it is an important building block used by cells to make
other lipids. Free fatty acids are incorporated into a large number of lipid
molecules essential to cellular structures such as the cell membrane.
Cholesterol
and fatty acids are not very soluble in water. In addition, unsaturated
fatty acids are susceptible to oxidative degradation and free radical
generation, which is harmful to cultured cells. Sigma offers a number of
formulations of cholesterol and free fatty acids that are designed to increase
their solubility and to protect them from oxidation.
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| Product # |
Product Name |
Description |
Application |
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C 9913 |
Cholesterol, synthetic |
- Liquid;
- Purity, not less than 95%;
- Synthesized from materials of non-animal origin;
- Molecular Weight=386.7.
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Use as a medium supplement in situations where
the use of animal-derived components is to be avoided.
Sigma is the only
commercial manufacturer of synthetic cholesterol.
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C 4951 |
Cholesterol, Water Soluble |
- Powder;
- Contains approx. 40 mg of cholesterol per gram; balance is
methyl-beta-cyclodextrin;
- Soluble at 200 mg/ml in water;
- Cell culture tested.
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Cholesterol is insoluble in water. This product
is formulated with methyl-beta-cyclodextrin to improve its solubility and make it more useful as a medium supplement.
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F 7050 |
Fatty Acid Supplement |
- Liquid;
- Proprietary mixture of fatty acids;
- Use at 0.25 to 0.5 ml per liter of medium;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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This mixture of fatty acids was developed
to optimize the growth of and expression of protein by Chinese Hamster Ovary (CHO) cells in serum-free culture. However, it may prove
useful for proliferation of a wide range of mammalian cell types.
Animal component-free.
May be used as a medium supplement in situations where
the use of animal-derived components is to be avoided.
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F 7175 |
Fatty Acid-Albumin
Supplement |
- Liquid;
- Contains 100 mg/ml of bovine serum albumin in DPBS (D 5652);
- Contains 2 moles of linoleic and 1 mole of oleic acid per mole of
albumin;
- Use at 0.5 to 1.0 ml per liter of medium;
- Sterile filtered;
- Cell culture tested, CHO, L929, and BHK.
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Useful for the development of serum-free
medium. Linoleic and oleic acids are important fatty acid found associated with albumins in nature. Albumin helps to improve the solubility of
fatty acids and to protect them from oxidation. Historically, fatty acids
have been added to cell cultures as components of serum.
This supplement is recommended for use with epithelial-derived cells. However, it may prove useful for proliferation of a wide range of mammalian
cell types.
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L 8384 |
Linoleic Acid-Albumin. |
- Powder, lyophilized;
- Contains 1.1 to 2.3 moles of linoleic acid per mole of bovine serum
albumin;
- Sterilized by gamma irradiation;
- Cell culture tested.
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Useful for the development of serum-free
medium. Linoleic acid is an important unsaturated fatty acid that is found associated with albumins in nature. Albumin helps to improve the solubility of
linoleic acid and to protect it from oxidation. Historically, it has been added to cell cultures
as a component of serum.
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L 9655 |
Linoleic,
Oleic, Acid-Albumin. |
- Liquid;
- Contains 100 mg/ml of bovine serum albumin in DPBS (D 5652);
- Contains 2 moles each of linoleic and oleic acids per mole of
albumin;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Useful for the development of serum-free
medium. Linoleic and oleic acids are important unsaturated fatty acids that are found associated with albumins in nature. Albumin helps to improve the solubility of
linoleic and oleic acids and to protect them from oxidation. Historically, they
have been added to cell cultures as a components of serum.
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L 0163 |
Linoleic,
Oleic, Arachidonic Acid-Albumin. |
- Liquid;
- Contains 100 mg/ml of bovine serum albumin in DPBS (D 5652);
- Contains 2 moles each of linoleic, oleic and arachidonic acids per mole of
albumin;
- Use at 0.5 to 1.0 ml per liter of medium;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Useful for the development of serum-free
medium. Linoleic, oleic and arachidonic acids are important fatty acids that are
found associated with albumins in nature. Albumin helps to improve the solubility of
linoleic, oleic and arachidonic acids and to protect them from oxidation. Historically,
they have been added to cell cultures as a components of serum.
When evaluating our
lipid loaded albumins, we recommend that L 0163 be evaluated first. It
is the most extensively lipid loaded albumin that Sigma offers.
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L 0288 |
Lipid Mixture 1, Chemically Defined. |
- Liquid;
- Contains non-animal derived fatty acids;
- Contains 2 ug/ml arachidonic and 10 ug/ml each linoleic, linolenic,
myristic, oleic, palmitic and stearic acids; 0.22 mg/ml cholesterol from
New Zealand sheep's wool, 2.2 mg/ml Tween-80, 70 ug/ml tocopherol acetate
and 100 mg/ml Pluronic F-68 solubilized in cell culture grade water (W
3500);
- Use at 1 to 10 ml per liter of medium;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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This supplement formulation is designed for use
in serum-free formulations.
It was developed to provide a source of fatty acids that are historically delivered into the cell culture system as
components of serum.
Tocopherol, in addition to being an important vitamin for cell proliferation, helps to reduce oxidation of the unsaturated
fatty acids.
Tween-80 increases the
solubility of the fatty acids and Pluronic F-68 helps to protect fragile cell
membranes from disruption during agitation of suspension cultures.
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O 3008 |
Oleic Acid-Albumin |
- Liquid;
- Contains 100 mg/ml of bovine serum albumin in DPBS (D 5652);
- Contains 2 moles of oleic acid per mole of
albumin;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Useful for the development of serum-free
medium. Oleic acid is an important fatty acid that is found associated with albumins in nature. Albumin helps to improve the solubility of
oleic acid and to protect it from oxidation. Historically, it has been added to cell cultures
as a component of serum.
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Sigma
offers a wide range of cell culture supplements. This table
is a listing of Sigma products that may be particularly useful in various biotechnology cell
culture application and techniques.
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| Product # |
Product Name |
Description |
Application |
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A 1345 |
Anti-oxidant Supplement (1000x) |
- Liquid;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Proprietary medium supplement specially formulated to boost cell growth for enhanced protein expression. May be used in combination with Polyamine Supplement (P 8483)
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C 6295 |
Cell Freezing Medium-DMSO Serum-free |
- Liquid, frozen;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Complete ready-to-use medium containing Minimum Essential Medium, dimethyl sulfoxide (8.7%) and methylcellulose.
Prepared according to the formulation of Waymouth (TCA Manual 2(1), 311-313 (1976).
Use to cryo-preserve cells.
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D 8537 |
Dulbecco's Phosphate Buffered Saline Modified |
- Liquid;
- Without calcium chloride and magnesium chloride;
- Cell culture tested;
- Sterile-filtered;
- Endotoxin tested.
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Calcium
and magnesium promote cell adhesion. Use this formulation of D-PBS when tissue
or cell dissociation or release is the objective.
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D 8662 |
Dulbecco's Phosphate Buffered Saline |
- Liquid;
- With calcium chloride and magnesium chloride;
- Cell culture tested;
- Sterile-filtered;
- Endotoxin tested;
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Known
as DPBS, this is a general use, isotonic saline solution for washing cells
and tissues. It
contains calcium and magnesium ions which facilitate cell attachment. |
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G 7513 |
L-Glutamine Solution, 200 mM |
- Liquid;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Prepared in cell culture grade water (W 3500). Useful for supplementing CHO, hybridoma
cell and other mammalian cell media.
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H 9394 |
Hanks'
Balanced Salt Solution Modified |
- Liquid;
- With sodium bicarbonate;
- Without calcium chloride, magnesium sulfate;
- Sterile-filtered;
- Endotoxin tested;
- Cell culture tested.
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Same
as H 9269, except that calcium and magnesium have been removed.
Calcium and magnesium
support cell adhesion.
Recommended for washing
adherent cells during passage.
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P 1300 |
Pluronic F-68, Non-ionic Detergent |
- Liquid;
- Cell culture tested.
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A media additive often used to protect cells growing in suspended culture systems from damage which may occur during
agitation and aeration.
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P 8483 |
Polyamine Supplement (1000x) |
- Liquid;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Proprietary
medium supplement formulated to boost cell growth for enhancement of protein expression.
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T 3924 |
Trypsin-EDTA Solution (1x) |
- Liquid, frozen;
- Contains 0.5 grams of porcine trypsin and 0.2 grams of EDTA-4Na per liter of Hank's Balanced
Salt Solution with phenol red;
- Trypsin powder is negative for parvovirus by 9 CFR;
- Sterile filtered;
- Cell culture tested.
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Use to release adherent cells from supporting surfaces during passage.
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T 6414 |
Trypsin Inhibitor Solution (1x) |
- Liquid, frozen;
- Contains chromatographically purified soybean trypsin inhibitor and iron supplemented calf
serum (C 8056);
- Prepared in Dulbecco's Phosphate Buffered Saline without calcium or
magnesium;
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
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Use to arrest the enzymatic activity of trypsin during passage of adherent cell lines.
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W 3500 |
Water
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- Liquid;
- Sterile filtered;
- Endotoxin tested, not more than 1.0 EU/ml;
- Cell culture tested.
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This water
is produced by a validated WFI system and is quality controlled for use in cell culture. Use cell culture grade water to
solubilize components and supplements for addition to cell culture medium.
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