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lipofectamine ltx with plustm reagent transfection reagent  (Thermo Fisher)


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    Thermo Fisher lipofectamine ltx with plustm reagent transfection reagent
    Lipofectamine Ltx With Plustm Reagent Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lipofectamine%C2%AE+ltx+reagent+with+plustm+reagent/lipofectamine+ltx+with+plustm+reagent+transfection+reagent/pmc12147355-44-45-56
    Average 90 stars, based on 1 article reviews
    lipofectamine ltx with plustm reagent transfection reagent - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Expressing:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Purification:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Transfection:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Article Title: FABP7 Regulates Acetyl-CoA Metabolism Through the Interaction with ACLY in the Nucleus of Astrocytes.
    Article Snippet: .. The constructed vectors were transfected into the cell line using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific Inc.) following the manufacturer’s instructions. ..

    Incubation:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Modification:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Selection:

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR2 antibody compositions and related methods
    Article Snippet: In short, the CHO cell supernatants were harvested by centrifugation and sterile filtered (0.2 μm) before FPLC-based affinity purification using Amsphere protein A columns (JSR Life Sciences) performed as described below. .. In-house expression and purification: pCB14-based expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/l) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/nil poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Expression vectors: Antibody sequences determined above to bind hROR2 were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain. .. Expression and purification of anti-ROR2 antibodies: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/L) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich. ..

    Article Title: Anthracycline-based antibody drug conjugates having high in vivo tolerability
    Article Snippet: Expression vectors: Fab sequences determined above to bind human CS1 were codon-optimized for human expression; variable domains were synthesized as DNA by GenScript (Piscataway, USA) and included within an expression vector containing suitable restriction sites and the appropriate constant domain (as per Waldmeier et al. 2016 for expression in HEK293T cells, and as per Beerli et al. 2015 for expression in CHO cells). .. HEK Expression and Purification: Expression vectors were transfected into HEK293T cells using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific, Reinach, Switzerland, 15388100); following a 1-day incubation (37° C., 5% CO2, growth media: Dulbecco's Modified Eagle Medium (DMEM) High Glucose (4.5 g/1) with L-Glutamine with 10% (v/v) Fetal Calf Serum (FCS), 100 IU/mL of Pen-Strep-Fungizone and 2 mM L-glutamine (all Bioconcept, Allschwil, Switzerland)), cells were expanded under selection conditions (2 μg/mL of puromycin (Sigma-Aldrich, Buchs SG, Switzerland, P8833-25 mg stock at 2 mg/mL)). .. Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 hrs at 37° C. and washed twice with PBS.

    Luciferase:

    Article Title: Bile Acids Modulate Colonic MAdCAM-1 Expression in a Murine Model of Combined Cholestasis and Colitis
    Article Snippet: Bile acids (Ursodeoxycholic acid, UDCA; Cholic acid, CA; Chenodeoxycholic acid, CDCA, Deoxycholic acid, DCA; and Lithocholic acid, LCA; Sigma-Aldrich) in DMSO were diluted 1:1,000 in culture media to a working concentration of 100 μM. .. For NF-κB luciferase reporter assays, lipofectamine® LTX Reagent with PLUSTM Reagent (ThermoFisher) was used to transfect the pNF-κB cis-reporter plasmid (Stratagene, La Jolla, CA, Cat. #219078-51) into TSECs, following manufacturer directions. .. NF-κB luciferase levels were measured using the Dual-Luciferase® Reporter Assay System (Promega, Madison, WI), according to manufacture directions.

    Plasmid Preparation:

    Article Title: Bile Acids Modulate Colonic MAdCAM-1 Expression in a Murine Model of Combined Cholestasis and Colitis
    Article Snippet: Bile acids (Ursodeoxycholic acid, UDCA; Cholic acid, CA; Chenodeoxycholic acid, CDCA, Deoxycholic acid, DCA; and Lithocholic acid, LCA; Sigma-Aldrich) in DMSO were diluted 1:1,000 in culture media to a working concentration of 100 μM. .. For NF-κB luciferase reporter assays, lipofectamine® LTX Reagent with PLUSTM Reagent (ThermoFisher) was used to transfect the pNF-κB cis-reporter plasmid (Stratagene, La Jolla, CA, Cat. #219078-51) into TSECs, following manufacturer directions. .. NF-κB luciferase levels were measured using the Dual-Luciferase® Reporter Assay System (Promega, Madison, WI), according to manufacture directions.

    other:

    Article Title: Human antibodies binding to ROR2
    Article Snippet: Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Article Title: ROR1 antibody compositions and related methods
    Article Snippet: Cells were split and further expanded (37° C., 5% CO2); once confluency was reached, tissue culture dishes were coated with 20 μg/ml poly-L-Lysine (Sigma-Aldrich, P1524) for 2 h at 37° C. and washed twice with PBS.

    Construct:

    Article Title: FABP7 Regulates Acetyl-CoA Metabolism Through the Interaction with ACLY in the Nucleus of Astrocytes.
    Article Snippet: .. The constructed vectors were transfected into the cell line using Lipofectamine® LTX Reagent with PLUSTM Reagent (Thermo Fisher Scientific Inc.) following the manufacturer’s instructions. ..



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    Image Search Results


    a, b The mRNA expression level of KLK7 in H4 cells treated with 30 µM memantine (Mem) (n=6, Student’s t -test, p=0.003) (a) , 100 µM NMDA, and 100 µM L-glutamate (L-Glu) (n=3, Dunnett’s test, p Ctrl vs NMDA <0.0001, p Ctrl vs L-Glu <0.0001) (b) , respectively. c Schematic representation of Aβ degradation assay in H4 cells. Synthetic Aβ40 was added to the conditioned medium of H4 cells treated with memantine. After 24 hours, remaining Aβ40 in conditioned medium was detected by immunoblotting. d The Aβ-degrading activity of H4 cells treated with 30 µM memantine. A representative immunoblot and quantified results are shown (Input=1.0, n=3, Student’s t -test, p=0.0205). e Cell viability of H4 cells treated with 30 µM memantine (n=3, Student’s t -test, p=0.3927). All data are shown as mean±SEM. *p<0.05, **p<0.01, ****p<0.0001; n.s., non-significant.

    Journal: bioRxiv

    Article Title: Inhibition of NF-κB signaling pathway in astrocytes facilitates amyloid-β clearance by kallikrein-related peptidase 7

    doi: 10.1101/2025.03.02.641088

    Figure Lengend Snippet: a, b The mRNA expression level of KLK7 in H4 cells treated with 30 µM memantine (Mem) (n=6, Student’s t -test, p=0.003) (a) , 100 µM NMDA, and 100 µM L-glutamate (L-Glu) (n=3, Dunnett’s test, p Ctrl vs NMDA <0.0001, p Ctrl vs L-Glu <0.0001) (b) , respectively. c Schematic representation of Aβ degradation assay in H4 cells. Synthetic Aβ40 was added to the conditioned medium of H4 cells treated with memantine. After 24 hours, remaining Aβ40 in conditioned medium was detected by immunoblotting. d The Aβ-degrading activity of H4 cells treated with 30 µM memantine. A representative immunoblot and quantified results are shown (Input=1.0, n=3, Student’s t -test, p=0.0205). e Cell viability of H4 cells treated with 30 µM memantine (n=3, Student’s t -test, p=0.3927). All data are shown as mean±SEM. *p<0.05, **p<0.01, ****p<0.0001; n.s., non-significant.

    Article Snippet: For transient expression of the luciferase constructs in H4 cells, Lipofectamine ® LTX Reagent & PlusTM Reagent (Thermo Fisher Scientific, catalog #15338100) was used following the manufacturer’s protocol.

    Techniques: Expressing, Degradation Assay, Western Blot, Activity Assay

    a Schematic representation of the establishment of luciferase constructs (Empty and h238 WT) and H4 stable cell lines. b, c Luciferase activity of H4 h238 WT treated with 100 µM NMDA, 100 µM L-glutamate (n=4, Dunnett’s test, p Ctrl vs NMDA =0.0038, p Ctrl vs L-Glu =0.0057) (b) , and 30 µM memantine (n=5, Student’s t -test, p=0.0003) (c) , respectively. d Schematic representation of luciferase constructs containing a deletion mutant and a motif mutant of the κB motif in the 238-bp KLK7 promoter region (h238 ΔκB motif and h238 mut-κB motif). e Luciferase activity of each luciferase construct transiently expressed in H4 cells (n=4, Dunnett’s test, p h238 WT vs h238 ΔκB motif =0.0432, p h238 WT vs h238 mut-κB motif =0.0318). f, g Luciferase activity of H4 h238 ΔκB motif (n=4, Dunnett’s test, p Ctrl vs NMDA =0.4847, p Ctrl vs L-Glu =0.3969) (f) and H4 h238 mut-κB motif (n=4, Dunnett’s test, p Ctrl vs NMDA =0.3716, p Ctrl vs L-Glu =0.5012) (g) treated with 100 µM NMDA or 100 µM L-glutamate. h, i Luciferase activity of H4 h238 ΔκB motif (n=5, Student’s t -test, p=0.8164) (h) and H4 h238 mut-κB motif (n=5, Student’s t -test, p=0.5011) (i) treated with 30 µM memantine. All data are shown as mean±SEM. *p<0.05, **p<0.01, ***p<0.001; n.s., non-significant.

    Journal: bioRxiv

    Article Title: Inhibition of NF-κB signaling pathway in astrocytes facilitates amyloid-β clearance by kallikrein-related peptidase 7

    doi: 10.1101/2025.03.02.641088

    Figure Lengend Snippet: a Schematic representation of the establishment of luciferase constructs (Empty and h238 WT) and H4 stable cell lines. b, c Luciferase activity of H4 h238 WT treated with 100 µM NMDA, 100 µM L-glutamate (n=4, Dunnett’s test, p Ctrl vs NMDA =0.0038, p Ctrl vs L-Glu =0.0057) (b) , and 30 µM memantine (n=5, Student’s t -test, p=0.0003) (c) , respectively. d Schematic representation of luciferase constructs containing a deletion mutant and a motif mutant of the κB motif in the 238-bp KLK7 promoter region (h238 ΔκB motif and h238 mut-κB motif). e Luciferase activity of each luciferase construct transiently expressed in H4 cells (n=4, Dunnett’s test, p h238 WT vs h238 ΔκB motif =0.0432, p h238 WT vs h238 mut-κB motif =0.0318). f, g Luciferase activity of H4 h238 ΔκB motif (n=4, Dunnett’s test, p Ctrl vs NMDA =0.4847, p Ctrl vs L-Glu =0.3969) (f) and H4 h238 mut-κB motif (n=4, Dunnett’s test, p Ctrl vs NMDA =0.3716, p Ctrl vs L-Glu =0.5012) (g) treated with 100 µM NMDA or 100 µM L-glutamate. h, i Luciferase activity of H4 h238 ΔκB motif (n=5, Student’s t -test, p=0.8164) (h) and H4 h238 mut-κB motif (n=5, Student’s t -test, p=0.5011) (i) treated with 30 µM memantine. All data are shown as mean±SEM. *p<0.05, **p<0.01, ***p<0.001; n.s., non-significant.

    Article Snippet: For transient expression of the luciferase constructs in H4 cells, Lipofectamine ® LTX Reagent & PlusTM Reagent (Thermo Fisher Scientific, catalog #15338100) was used following the manufacturer’s protocol.

    Techniques: Luciferase, Construct, Stable Transfection, Activity Assay, Mutagenesis

    a Schematic representation of the canonical NF-κB activation pathway with the targets of the inhibitors IKK-16 and JSH-23 indicated. b The mRNA expression level of KLK7 in H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23 (n=6, Student’s t -test, p Ctrl vs IKK-16 =0.0270, p Ctrl vs JSH-23 =0.0103). c The Aβ-degrading activity of H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23. Representative immunoblots and quantified results are shown (Input=1.0, n=4, Student’s t -test, p Ctrl vs IKK-16 =0.0010, p Ctrl vs JSH-23 =0.0082). d Cell viability of H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23 (n=4, Student’s t -test, p Ctrl vs IKK-16 =0.9618, p Ctrl vs JSH-23 =0.9043). e Luciferase activity of H4 h238 WT treated with 2 µM IKK-16 or 30 µM JSH-23 (n=4 (IKK-16), n=6 (JSH-23), Student’s t -test, p Ctrl vs IKK-16 <0.0001, p Ctrl vs JSH-23 <0.0001). All data are shown as mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; n.s., non-significant.

    Journal: bioRxiv

    Article Title: Inhibition of NF-κB signaling pathway in astrocytes facilitates amyloid-β clearance by kallikrein-related peptidase 7

    doi: 10.1101/2025.03.02.641088

    Figure Lengend Snippet: a Schematic representation of the canonical NF-κB activation pathway with the targets of the inhibitors IKK-16 and JSH-23 indicated. b The mRNA expression level of KLK7 in H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23 (n=6, Student’s t -test, p Ctrl vs IKK-16 =0.0270, p Ctrl vs JSH-23 =0.0103). c The Aβ-degrading activity of H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23. Representative immunoblots and quantified results are shown (Input=1.0, n=4, Student’s t -test, p Ctrl vs IKK-16 =0.0010, p Ctrl vs JSH-23 =0.0082). d Cell viability of H4 cells treated with 2 µM IKK-16 or 30 µM JSH-23 (n=4, Student’s t -test, p Ctrl vs IKK-16 =0.9618, p Ctrl vs JSH-23 =0.9043). e Luciferase activity of H4 h238 WT treated with 2 µM IKK-16 or 30 µM JSH-23 (n=4 (IKK-16), n=6 (JSH-23), Student’s t -test, p Ctrl vs IKK-16 <0.0001, p Ctrl vs JSH-23 <0.0001). All data are shown as mean±SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001; n.s., non-significant.

    Article Snippet: For transient expression of the luciferase constructs in H4 cells, Lipofectamine ® LTX Reagent & PlusTM Reagent (Thermo Fisher Scientific, catalog #15338100) was used following the manufacturer’s protocol.

    Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Luciferase