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Complete amino acid mix

Manufactured by Promega
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The Complete Amino Acid Mix is a laboratory product that provides a comprehensive set of essential and non-essential amino acids. It is a balanced mixture of the 20 standard amino acids that are the basic building blocks of proteins. The mix is intended for use in various research and analytical applications that require a complete amino acid solution.

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Lab products found in correlation

Rnasin ribonuclease inhibitor, by Promega (1 mentions) Tnt quick coupled transcription translation system, by Promega (1 mentions) Rneasy minelute cleanup kit, by Qiagen (1 mentions) Qiaquick gel extraction kit, by Qiagen (1 mentions) Envision multilabel plate reader, by PerkinElmer (1 mentions) Protector rnase inhibitor, by Roche (1 mentions) Mmessage mmachine t7 transcription kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Amino acids (6 citations) Amino acid mix (3 citations)

2 protocols using complete amino acid mix

1

In Vitro Protein Translation Optimization

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In vitro translation was carried out using TnT Quick Coupled Transcription/Translation System (L1170, Promega) following the manufacturer’s protocol. Briefly, TnT T7 Quick Master Mix was thawed quickly by hand, whereas the other components were thawed on ice. A reaction mixture (50 μl) was prepared. TnT T7 Master Mix (25 μl) was mixed with 2 μl of TnT Reaction Buffer, 1 μl of TnT RNA Polymerase, 2 μl of Complete Amino Acid Mix (L4461, Promega), 1 μl of RNasin Ribonuclease Inhibitor (N2511, Promega), 2 μl of DNA plasmid, and 17 μl of nuclease-free water. The reaction mixture was mixed thoroughly and incubated at 30°C for 90 min. Immunoblotting was performed to confirm successful translation.
Yuen K.C., Slaughter B.D, & Gerton J.L. (2017). Condensin II is anchored by TFIIIC and H3K4me3 in the mammalian genome and supports the expression of active dense gene clusters. Science Advances, 3(6), e1700191.
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2

In vitro translation and folding of GFP-Gαi1

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A pCI-neo plasmid expressing rat Gαi1 with an internal
GFP tag between amino acids 122 and 123 of rat Gαi1 with the
amino acid sequence SGGGGS as a linker at the N- and C-terminal ends of GFP was
obtained from A. V. Smrcka (University of Michigan). The GFP-tagged
Gαi1 pCI-neo plasmid was linearized by digestion with Cla
I and purified using a QIAquick Gel Extraction Kit (Qiagen) for use as an in
vitro transcription template. Capped GFP-Gαi1 mRNA transcripts
were produced using the mMESSAGE/ mMACHINE T7 Transcription Kit (Life
Technologies). We purified mRNA with an RNeasy MinElute CleanUp Kit (Qiagen).
The mRNA was diluted to 500 μg/ml in water. GFP-Gαi1mRNA (500 ng) was translated, and protein folding ensued in 25-ml reactions
containing 12.5 μl of micrococcal nuclease–treated WGE (Promega),
3.5 μl of complete amino acid mix (Promega), 20 mM potassium acetate, 0.5
μl of Protector RNase Inhibitor (Roche), and purified Ric-8 proteins (10
nM to 2.5 μM) for 5-hour kinetic reactions at 25°C in Nunc
384-well black flat bottom plates. GFP fluorescence (485-nm excitation/ 535-nm
emission) was measured using an EnVision Multilabel Plate Reader
(PerkinElmer).
Papasergi-Scott M.M., Stoveken H.M., MacConnachie L., Chan P.Y., Gabay M., Wong D., Freeman R.S., Beg A.A, & Tall G.G. (2018). Dual phosphorylation of Ric-8A enhances its ability to mediate G protein α subunit folding and to stimulate guanine nucleotide exchange. Science signaling, 11(532), eaap8113.
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