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T2000-01 TC-100 Insect Medium w/ L-Glutamine, w/o NaHCO3 (Powder)

Specifications
References
Grade
Cell Culture Grade
EU Commodity Code
38210000
Shipping Temp
RT
Storage Temp
RT/4°C

TC-100 is a modification done by Dr. Gardiner and Dr. Stockdale of the original formulation of Grace's insect medium. Sucrose, fructose and several Kreb's cycle intermediates were omitted from the original formula. The modification was done to optimize the production of Autographia californica NPV virions by cells from the fall armyworm, Spodoptera frugiperda.

TC-100 is a fully defined medium and does not require supplementation with insect hemolymph. Instead, tryptose broth and fetal bovine serum provide the necessary growth factors. This medium supports the growth of several lepidopteran cell lines including Spodoptera frugiperda. It is specifically used for production of baculoviruses in lepidopteran cell lines. Users are advised to review the literature for recommendations regarding medium supplementation and physiological growth requirements specific for different cell lines.
Appearance
White to off white, homogenous, free flowing powder
Solubility
Colorless to light yellow, clear, complete
pH
As Reported
Directions per Liter
Dissolve 19.07g in 800-900ml of ddH2O stirring gently until completely solubilized. If required, add sodium bicarbonate. Adjust pH of the medium to the desired level. Add additional water to bring the solution to1L Filter-sterilize using a 0.22 micron membrane filter. Aliquot into sterile containers. Do not autoclave. Contains heat-labile compounds that can be damaged with autoclaving.
Note: On occasion, this formulation seems reluctant to dissolve. If continuous stirring with warmth has not produced complete dissolution, add 1.0N HCl slowly until solution turns crystal clear with light yellowish tone (pH 4.2-4.5). Let solution cool to RT, then back titrate with 1.0N NaOH to pH 6.2-6.4. Components will remain solubilized.
Storage and Stability
Store powdered media at RT. Stable for 12 months after receipt. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept at 4°C and used within a short period of time.
Media Formulation
Components shown in g/L
Inorganic Salts
Calcium Chloride0.9966
Magnesium Chloride1.068189
MagnesiumSulfate1.357858
Potassium Chloride2.87
Sodium BicarbonateAbsent
Sodium Phosphate Monobasic0.876923
Amino Acids
L-Alanine0.225
L-Arginine•HCl0.70
L-Asparagine0.35
L-Aspartic Acid0.35
L-Cystine•2HCl0.025
L-Glutamic Acid0.60
L-Glutamine0.60
Glycine0.65
L-Histidine2.50
L-Isoleucine0.05
L-Leucine0.075
L-Lysine•HCl0.625
L-Methionine0.05
L-Phenylalanine0.15
L-Proline0.35
L-Serine0.55
L-Threonine0.175
L-Tryptophan0.10
L-Tyrosine•2Na•2H2O0.07202
L-Valine0.10
Vitamins
p-Aminobenzoic Acid0.00002
D-Biotin0.00001
Choline Chloride0.0002
FolicAcid0.00002
Myo-Inositol0.00002
Niacin0.00002
D-Pantothenic Acid•Ca0.00002
Pyridoxine•HCl0.00002
Riboflavin0.00002
Thiamine•HCl0.00002
Other
D(+)-Glucose1.00
Tryptose Broth2.60
References
1. Wang L., et al. 2018. Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) Induces Host Cell Autophagy to Benefit Infection. Viruses. 10(1), 14. doi:10.3390/v10010014. 2. Dong ZQ, et al. Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells. 2017. Sci Rep. 7:46187. doi: 10.1038/srep46187. 3. Gorbenko, O. et al., Monoclonal antibodies with selective specificity towards different glycosylation isoforms of FGFR1.2009) Hybridoma (Larchmt). 28(4):287-293.
USBio References
1. Wang L., et al. 2018. Bombyx mori Nuclear Polyhedrosis Virus (BmNPV) Induces Host Cell Autophagy to Benefit Infection. Viruses. 10(1), 14. doi:10.3390/v10010014|2. Dong ZQ, et al. Baculovirus LEF-11 Hijack Host ATPase ATAD3A to Promote Virus Multiplication in Bombyx mori cells. 2017. Sci Rep. 7:46187. doi: 10.1038/srep46187.|3. Gorbenko, O. et al., Monoclonal antibodies with selective specificity towards different glycosylation isoforms of FGFR1.2009) Hybridoma (Larchmt). 28(4):287-293.
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