TfR-sfGFP-myc tag-SpyCatcher variants were expressed in suspension Expi293 cells (Thermo Fisher) cultured in Expi293 Expression media (Thermo Fisher) supplemented with 50 U/mL penicillin/streptomycin (Thermo Fisher). Cells were grown in a humidified Multitron Cell incubator (Infors HT) at 37 °C with 7% CO2, rotating at 110 to 125 rpm. Cells at a density of 3.0 × 106 cells/mL were transfected with 2.7 μL ExpiFectamine 293 Reagent per 1 μg of plasmid DNA. ExpiFectamine transfection enhancers (Thermo Fisher) were added 16 to 22 h after transfection. Cells were grown for 48 h and then analyzed.
Expifectamine transfection enhancers
Manufactured by Thermo Fisher Scientific
ExpiFectamine transfection enhancers are a family of cationic lipid-based transfection reagents designed to enhance the efficiency of transient protein expression in mammalian cells. These reagents are formulated to facilitate the delivery of DNA, RNA, or other macromolecules into the cells for various research applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using expifectamine transfection enhancers
1
Expressing TfR-sfGFP-myc tag-SpyCatcher variants
2
Protein Complex Assembly in Expi293F Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Three component expressions
were carried out with either (i) TfR-sfGFP-SnoopTag2 plus HA-MBP-SnoopLigase2
N775Q-KDEL plus RBD-DogTag2-SpyTag or (ii) TfR-sfGFP-SnoopTag2 plus
HA-MBP-SnoopLigase2 N775Q-KDEL plus RBD-DogTag2 NA-SpyTag in Expi293F
cells (Thermo Fisher) cultured in Expi293 expression media (Thermo
Fisher). Cells at a density of 3.0 × 106 cells/mL
that had previously been growing in a medium supplemented with 50
U/mL penicillin/streptomycin (Thermo Fisher) were transfected in Expi293
expression media with no antibiotics present. Plasmids were transiently
transfected with 2.7 μL of ExpiFectamine 293 Reagent per 1 μg
of plasmid DNA with equal amounts of each plasmid added in each of
the three component transfections. A mock transfection where the plasmid
DNA was omitted was also carried out in parallel. Cells were grown
in a humidified Multitron Cell incubator (Infors HT) at 37 °C
with 7% (v/v) CO2, rotating at 110–125 rpm. ExpiFectamine
transfection enhancers (Thermo Fisher) were added 16–22 h after
transfection. Cells were grown for 4 days and then analyzed.
were carried out with either (i) TfR-sfGFP-SnoopTag2 plus HA-MBP-SnoopLigase2
N775Q-KDEL plus RBD-DogTag2-SpyTag or (ii) TfR-sfGFP-SnoopTag2 plus
HA-MBP-SnoopLigase2 N775Q-KDEL plus RBD-DogTag2 NA-SpyTag in Expi293F
cells (Thermo Fisher) cultured in Expi293 expression media (Thermo
Fisher). Cells at a density of 3.0 × 106 cells/mL
that had previously been growing in a medium supplemented with 50
U/mL penicillin/streptomycin (Thermo Fisher) were transfected in Expi293
expression media with no antibiotics present. Plasmids were transiently
transfected with 2.7 μL of ExpiFectamine 293 Reagent per 1 μg
of plasmid DNA with equal amounts of each plasmid added in each of
the three component transfections. A mock transfection where the plasmid
DNA was omitted was also carried out in parallel. Cells were grown
in a humidified Multitron Cell incubator (Infors HT) at 37 °C
with 7% (v/v) CO2, rotating at 110–125 rpm. ExpiFectamine
transfection enhancers (Thermo Fisher) were added 16–22 h after
transfection. Cells were grown for 4 days and then analyzed.
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