Full length HA-IKKβ protein was obtained from in-vitro translation kit (catalog no. L1170; Promega). BL21 E. coli transformed with empty vector or pGEX-GST-ARID1A-F2 truncated plasmid was induced by Isopropyl-1-thio-β-D-galactopyranoside (IPTG) (0.5 mM) at 16 °C for 5 h. After sonication, incubate the cell lysate with glutathione-Sepharose beads for 1 h at 4 °C. Recombinant proteins binding to beads were incubated with HA-IKKβ protein for 3 h at 4 °C. Beads were subsequently harvested through centrifugation and washed three times by 0.2% NP-40 buffer before boiled and loaded to SDS-PAGE. Full scan blots, see the Source Data file.
In vitro translation kit
Manufactured by Promega
Sourced in United States
The In-vitro translation kit is a laboratory tool designed to synthesize proteins from genetic material in a controlled, cell-free environment. The kit provides the necessary components, including ribosomes, tRNAs, and energy sources, to facilitate the translation of mRNA into functional proteins. This product enables researchers to study protein expression, characterization, and modifications without the complexity of a living cell system.
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Lab products found in correlation
2 protocols using in vitro translation kit
1
Protein Interaction Assay with HA-IKKβ
2
GST-Fusion Protein Interaction Analysis
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Glutathione-S-transferase (GST) or GST-fusion proteins were expressed and purified, according to the manufacturer’s instructions (Amersham Biosciences, England). In vitro-translated His-RACK1 was incubated with GST or GST-GNA14 bound to glutathione-Sepharose beads (Amersham Biosciences, England), and the adsorbed proteins were analyzed by immunoblotting analysis. In vitro-translation kit was purchased from Promega (Wisconsin, USA).
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