Expi293 Met(–)Expression Medium は、cGMPで製造され、化学的に定義され、血清不含、タンパク質不含、動物由来成分不含、メチオニン不含の、Expi293F細胞増殖およびトランスフェクション用の培地です。この培地は、哺乳類HEK293細胞から得られた迅速で高収量の一過性タンパク質発現を実現するExpi293プラットフォームの一部です。タンパク質中のメチオニン残基の標識化が要求されるアプリケーション用でExpiFectamine 293 Met (–) Transfection Kitと組み合わせて使用するように設計されています。注:この培地は、メチオニンを添加しない限り、細胞増殖や発現をサポートしません。
Expi293 Met(–) Expression Medium の特徴は以下のとおりです。 •1.5 x 107細胞/mL以上の密度までの、懸濁液293培養物の増殖をサポートします。 • Expi293 Expression Mediumと同等の性能(カタログ番号 A1435101) •メチオニンを含みません。 •フェノールレッドを含みません。 •GlutaMAX I サプリメント配合 •メチオニンの追加が必要
1 L Expi293 Met(–) Expression Medium、2~8℃で保存してください(光から保護してください)。
よくあるご質問(FAQ)
I am performing L-selenomethionine labeling and getting very low protein yields. What could be going wrong?
L-selenomethionine is a known cell toxin. Therefore, it is important to grow cells in the present of this amino acid for the shortest amount of time required to achieve maximal labeling. We recommend adding L-selenomethionine to the culture just prior to transfection (in Expi293 Met (-) Expression Medium). You may also starve cells for 6 hr by culturing cells in Expi293 Met (-) Expression Medium (without supplementation) and then proceed with L-selenomethionine add-back and transfection.
I'm culturing my Expi293F cells in Expi293 Met (-) Expression Medium but they are not growing. What could be going wrong?
Expi293 Met (-) Expression Medium does not contain methionine, which is an essential nutrient for cell culture. Therefore, methionine supplementation is required. For methionine labeling, cells should be grown in Expi293 Met (-) Expression Medium and starved (i.e., grown in the absence of methionine supplementation) for 6 hr prior to addition of 13C-methyl-L-methionine, or similar methionine isotope.
How do I use Expi293 Met (-) Expression Medium for methionine protein labeling?
Expi293 Met (-) Expression Medium is not intended to support long-term cell culture as the media is deficient of methionine, an essential amino acid for cell survival. When you are ready to start your labeling experiment, transfer Expi293F Cells growing in Expi293 Expression Medium to Expi293 Met (-) Expression Medium by centrifuging the cell suspension (200 x g for 5 min) and resuspending the cell pellet in Expi293 Met (-) Expression Medium. Cells may be starved for 6 hr prior to supplementation with L-selenomethionine, isotopic-labeled methionine, or other derivatives.
What is the difference between Expi293 Expression Medium and Expi293 Met (-) Expression Medium?
The formulation of Expi293 Met (-) Expression Medium is identical to that of Expi293 Expression Medium except for the absence of methionine. Expi293 Met (-) Expression Medium allows for the addition of isotopic-labeled methionine or derivatives to promote labeling of proteins for structural analysis.