Beijing BioMed Gene Technology Co., Ltd. provided plasmid vector PET28a, E. coli BL21 (DE3). BCA protein quantitative kits and plasmid small extraction kits were purchased from Sangon Bioengineering (Shanghai) Co., Ltd.. Peptone, yeast powder and agar powder were purchased from Oxid Ltd., and casein was purchased from Beijing Solarbio Biotechnology Co., Ltd. Our laboratory synthesized colloidal gold particle. Sino Biological Inc. kindly donated rabbit monoclonal antibody against SARS -CoV-2 N protein. Isopropyl-β-d -thiogalactopyranoside(IPTG) and G25 dextran gel were purchased from GE Healthcare, and Ni NTA agarose resin, SP strong cation agarose resin, and CM weak cation agarose resin were purchased from Tosoh Bioscience.
Isopropyl β d thiogalactopyranoside iptg
Manufactured by GE Healthcare
Sourced in United States
Isopropyl-β-d-thiogalactopyranoside (IPTG) is a chemical compound used in molecular biology and microbiology laboratories. It is a synthetic analog of the natural lactose molecule and is commonly used as an inducer in the expression of recombinant proteins in bacterial systems, such as Escherichia coli. IPTG binds to the lac repressor, allowing transcription of the target gene to occur.
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Lab products found in correlation
2 protocols using isopropyl β d thiogalactopyranoside iptg
1
SARS-CoV-2 N Protein Purification Protocol
2
Expression and Structural Prediction of Adenovirus Hex Protein
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According to the sequence of CAdV-2 Hex (GenBank accession number AC000020.1), the primers were designed to amplify an 897-bp fragment (F: 5′-1654-TATGAATTC GACCTCCGAGTGGATGGG-1671-3′, R: 5′-2533-GAGCTCGAG GGAGTTAGAGTACAGCAG—2550-3′). The fragment was cloned into a pET30a (+) vector (Tiangen, Beijing, China) to construct the expression plasmids. The expression plasmid was transformed into E. coli BL21 competent cells (Tiangen); the truncated Hex protein was produced via isopropyl-β-D-thiogalactopyranoside (IPTG, GE Healthcare, Chicago, IL, USA) induction using pET30a- Hex E. coli at 37 °C. The truncated Hex protein was detected by SDS-PAGE and Western blotting (WB) analysis by anti-His mAb. Immunoreactive bands were verified by an enhanced chemiluminescence system (ECL; PerkinElmer Life Sciences, Fremont, CA, USA) [15 (link)]. The three-dimensional (3D) structure of the expressed protein was predicted with PyMOL software based on data from the SWISS-MODEL online server.
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