Flp-in t-rex
WebAs a first step to Flp-In T-rex cell lines, you have to create the so called host cell line, which is generated through stable random integration of two plasmids pFRT/lacZeo … WebDescription Once Flp-InT-REx host cell line containing integrated FRT site has been created, subsequent generation of Flp-InT-REx cell lines expressing gene (s) of interest is rapid and efficient Allows generation of isogenic, inducible stable cell lines Permits polyclonal selection of stable expression cell lines
Flp-in t-rex
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WebCulturing the Flp-In™ T-REx™-293 Cell Line, Continued Thawing Cells The following protocol is designed to help you thaw cells to initiate cell culture. The Flp-In™ T-REx™ … WebUpon isolation of DPCs by PxP, samples were analysed using western blotting using the indicated antibodies. d HeLa T-REx Flp-In SPRTN-ΔC cells were complemented with YFP, YFP-SPRTN-Strep variants as indicated (wildtype (WT), E112Q (EQ), R185A (ZBD*)) and treated as shown in Fig. , before DPCs were isolated using PxP and analysed by …
WebAs a first step to Flp-In T-rex cell lines, you have to create the so called host cell line, which is generated through stable random integration of two plasmids pFRT/lacZeo (introducing the FRT...
WebMar 22, 2024 · Further investigation in differentiated HuH-7 cells expressing NTCP and NTCP-overexpressing Flp-In T-REx 293 cells revealed that the mechanism of action of everolimus on NTCP is direct inhibition and mTOR-independent. Structural analogs of everolimus inhibited NTCP-mediated TCA uptake, however, functional analogs did not … WebApr 26, 2024 · Flp-In T-REx 293 cells expressing c-IBD-SBP-YAP1C were incubated in DPBS containing 10 mM 3-AT and supplemented or not with 1 mM H 2 O 2. After 10 min, free thiol groups were blocked with NEM, and the samples were processed as depicted in panel A and subsequently subjected to immunoblotting with rabbit pre-immune serum …
WebApr 11, 2024 · The Flp-In TM 293 T-REx TM pcDNA.5/FRT/TO inducible Tet-ON expression system was used to create stable, inducible cell lines, whereby the Ku70-BioID2 fusion protein (wild-type, Ku70 S155A, or Ku70 S155D) or the Ku70 NLS-BioID2 fusion protein was stably integrated in the T-REx 293 cell line and under the control of a doxycycline …
WebApr 1, 2024 · Protocol used to generate stable Flp-In T-REx-HEK 293 cell l ines expressing WT or . mutant GCase (E326K o r L444P) as. a V5-FLAG-tagged protein using a tetracycline-inducible system. citizen credit cooperative bank puneWebjor components of the Flp-In™ T-REX™ System include: • A Flp-In™ target site vector, pFRT/lacZeo, for generation of a Flp-In™ host cell line containing an integrated FRT site. … dichlophosWebMar 3, 2024 · Generation of Stable Inducible Cell Pools and BioID Labeling. Cell lines were generated in HEK293 Flp-In T-REx 293 (Invitrogen; used for BioID, affinity purification coupled to MS and some of the streptavidin-immunobot analyses) or HeLa Flp-In T-REx cells (used for BioID and the BioID-immunoblot analyses) grown at 37 °C in Dulbecco’s … citizen credit union online bankingWebFlp-in T-Rex HEK293 cells were maintained in the presence of 100 μg/mL zeocin and 15 μg/mL blasticidin. To generate stable doxycycline (dox)-inducible secreted proteins or FLAG–Cas9-expressing cells, plasmids encoding the respective proteins were co-transfected with pOG44, encoding the FLP recombinase, into Flp-in T-Rex HEK293 … citizen credit co-operative bank ltd. mumbaiWebAug 20, 2024 · The Flp-In system integrates a vector into cell lines at a specific genomic locus, but has not been used for integration of more than one gene. Here we present a modification of the Flp-In... citizen credit cooperative bank recruitmentWebApr 21, 2024 · Flp-In t -293细胞系包含稳定整合的pFRT lacZeo和pcDNA 6 TR (来自T-REx系统)。 用Flp- in表达载体和Flp重组酶载体pOG44共转染Flp- in细胞系,可将表达载体定向整合到每个细胞的同一位点,保证基因 … dichj how have you beenWebI am trying to generate stable cell line by using Flp-In 293 T-REx cells. I use 9:1 (w/w) ratio of pOG44:pcDNA-GOI and lipofectamine as a transfection reagent. After 48 hours transfection, I... citizen credit cooperative bank panjim