psPAX2 (Addgene, no.12260) was a gift from Didier Trono. pCDNA3.3_CoV2_B.1.1.7 (Addgene, no.170451) for Alpha-S and pcDNA3.3-SARS2-B.1.617.2 (Addgene, no.172320) for Delta-S proteins, were gifts from David Nemazee36 (link). pTwist-SARS-CoV-2 Δ18 B.1.351v1 (Addgene, no.169462) for Beta-S protein was a gift from Alejandro Balazs37 (link). Lentiviral vector, pWPI-ffLuc-P2A-EGFP for luciferase reporter assay and pTRC2puro-ACE2-P2A-TMPRSS2 for the generation of 293T cell line susceptible to SARS-CoV-2 infection was created from pWPI-IRES-Puro-Ak-ACE2-TMPRSS2, a gift from Sonja Best (Addgene, no.154987) by In-Fusion® technology (Takara Bio). pcDNA3.4 expression plasmids encoding SARS-CoV-2 S proteins with human codon optimization and 19 a.a deletion of C-terminus (C-del19) from Wuhan, D614G, and Omicron were generated by assembly of PCR products, annealed oligonucleotides, or artificial synthetic gene fragments (Integrated DNA Technologies, IDT) using In-Fusion® technology. For Delta plus, Kappa and Lambda variants, S proteins with only RBD, D614, and P681 mutations were created from pcDNA3.4 encoding human codon-optimized Wuhan S protein (C-del19). LentiX-293T cells (Takara Bio) and 293T cells were maintained in culture with Dulbecco’s Modified Eagle’s Medium (DMEM) containing 10% fetal bovine serum (FBS), penicillin-streptomycin (Nacalai tesque), and 25 mM HEPES (Nacalai tesque).
Hepes
Manufactured by Takara Bio
Sourced in China
HEPES is a chemical compound used as a buffer solution in biological research and cell culture applications. It helps maintain a stable pH environment for cells and biological samples. HEPES is commonly used in various laboratory procedures to ensure optimal conditions for cell growth and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
In fusion technology, by Takara Bio (1 mentions)
Lenti x 293t cells, by Takara Bio (1 mentions)
Penicillin streptomycin, by Takara Bio (1 mentions)
Pspax2, by Addgene (1 mentions)
Pcdna3 3 cov2 b 1, by Addgene (1 mentions)
Pcdna3 3 sars2 b 1, by Addgene (1 mentions)
Pwpi ires puro ak ace2 tmprss2, by Addgene (1 mentions)
Fetal bovine serum (fbs), by Takara Bio (1 mentions)
Penicillin, by Takara Bio (1 mentions)
Streptomycin, by Takara Bio (1 mentions)
Rpmi 1640 medium, by Takara Bio (1 mentions)
2 protocols using hepes
1
Expression Plasmids for SARS-CoV-2 Variants
2
Culturing the INS-1 Rat Insulinoma Cell Line
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The rat insulinoma cell line INS-1 was obtained from the China Center for Type Culture Collection (Wuhan University, Wuhan, China). RPMI-1640 cell culture medium and fetal bovine serum (FBS) were purchased from HyClone (GE Healthcare Life Sciences, Logan, UT, USA).
INS-1 cells were maintained in RPMI-1640 medium containing 11 mM glucose supplemented with 10% (v/v) heat-inactivated FBS, 100 U/ml penicillin, 100 U/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate and 50 µM β-mercaptoethanol (all Takara Biotechnology Co., Ltd., Dalian, China). Culture medium was replaced daily until cell confluency reached 80–90%.
INS-1 cells were maintained in RPMI-1640 medium containing 11 mM glucose supplemented with 10% (v/v) heat-inactivated FBS, 100 U/ml penicillin, 100 U/ml streptomycin, 10 mM HEPES, 1 mM sodium pyruvate and 50 µM β-mercaptoethanol (all Takara Biotechnology Co., Ltd., Dalian, China). Culture medium was replaced daily until cell confluency reached 80–90%.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!