96 well flat bottom plates (Thermo Fisher)

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96 Well Plate Individual

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Thermo Scientific Abgene polypropylene microplates and deepwell plates offer storage security for assays compound libraries or storing samples for either intermediate or long term use Abgene microtiter plates are manufactured to exacting specifications in our Class 100 000 clean room ISO 9001 conditions using high quality medical grade virgin polypropylene resins which ensure confidence in the quality and performance of the storage plates Designed to ANSI standards these microplates and deepwell plates are compatible with a variety of automated liquid handling for high throughput workflows The polypropylene storage plates are offered with spindle pyramidal U and V bottom wells along with a wide range of volumes to accommodate most applications

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ab0765

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None

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Laboratory Plastics and Supplies|Microplates, Dishes and Flasks

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Thermo Fisher 96 well flat bottom plates
Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in <t>96-well</t> flat-bottom plates in the presence

Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in <t>96-well</t> flat-bottom plates in the presence

96 well flat bottom plates - by Bioz Stars, 2021-01

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Images

1) Product Images from "Triptolide, a constituent of immunosuppressive Chinese herbal medicine, is a potent suppressor of dendritic-cell maturation and trafficking"

Article Title: Triptolide, a constituent of immunosuppressive Chinese herbal medicine, is a potent suppressor of dendritic-cell maturation and trafficking

Journal:

doi: 10.1182/blood-2005-03-0854

Figure Legend Snippet: Effect of TPT on the proliferative response of bidirectional MLR and anti-CD3/CD28–stimulated CD4 + T cells. Allogeneic human PBMCs from 2 different donors (3 × 10 5 cells/well each) were cocultured in 96-well flat-bottom plates in the presence

Techniques Used:

2) Product Images from "Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators"

Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators

Journal: Journal of Clinical Investigation

doi: 10.1172/JCI25901

Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.

Figure Legend Snippet: Nonsecosteroidal VDR ligands are potent agonists in keratinocytes and PBMCs. ( A ) LY2108491 and LY2109866 are potent inhibitors of keratinocyte proliferation. KerTr cells plated in 96-well plates were dosed with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 72 hours at 37°C before BrdU incorporation into DNA was analyzed as a measure of cell proliferation. Results (mean α SEM) of experiments performed in triplicate are shown. ( B ) Nonsecosteroidal VDR ligands are potent inducers of CYP24 gene expression in keratinocytes. TaqMan quantitative RT-PCR (Q-PCR) was performed on total RNA prepared from KerTr cells treated with various concentrations of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. Levels of GAPDH mRNA were measured in all the samples, and the results were normalized and presented as fold induction (α SEM) compared with normalized CYP24 levels in vehicle-treated cells. ( C ) Nonsecosteroidal VDR ligands are efficacious in TPA- and PHA-activated PBMCs. Primary cells isolated from donors were stimulated with TPA (100 ng/ml) and PHA (25 μl/ml) and treated with vehicle or 100 nM each of 1,25-(OH) 2 D 3 , LY2108491, or LY2109866 for 24 hours. TaqMan Q-PCR was performed on RNA obtained from vehicle-treated or VDR ligand–treated samples, using primer pairs and probes for IL-2, IL-4, IL-10, GATA3, and GAPDH. The amount of IL-2, IL-4, IL-10, and GATA3 transcripts relative to GAPDH transcripts is shown as mean α SEM of quadruplicate experimes.

Techniques Used: BrdU Incorporation Assay, Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Isolation

3) Product Images from "Effect of azithromycin on a red complex polymicrobial biofilm"

Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

Journal: Journal of Oral Microbiology

doi: 10.1080/20002297.2017.1339579

Effect of azithromycin up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Azithromycin at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h under anaerobic conditions. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

Figure Legend Snippet: Effect of azithromycin up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Azithromycin at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h under anaerobic conditions. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

Techniques Used: Incubation

Formation of mono- and polymicrobial biofilms in a 96-well plate model after 48 h of incubation at 37°C under anaerobic condition. Native bacterial growth with addition of uncultured growth medium and no antibiotic served as controls. Adherent biofilms were stained with 0.1% crystal violet and the optical density at AU 620 was measured. Data represent the mean AU 620 value of a minimum of three biological replicates.

Figure Legend Snippet: Formation of mono- and polymicrobial biofilms in a 96-well plate model after 48 h of incubation at 37°C under anaerobic condition. Native bacterial growth with addition of uncultured growth medium and no antibiotic served as controls. Adherent biofilms were stained with 0.1% crystal violet and the optical density at AU 620 was measured. Data represent the mean AU 620 value of a minimum of three biological replicates.

Techniques Used: Incubation, Staining

Effects of azithromycin and amoxicillin + metronidazole (1:1 ratio) up to 5.0 mg/L on formation polymicrobial biofilms after 48 h of anaerobic incubation at 37°C in a 96-well plate model. Azithromycin and amoxicillin + metronidazole (1:1 ratio) at concentrations 0–100 mg/L were incubated with bacterial cultures. Data points represent the mean AU 620 value of a minimum of three biological replicates and the standard deviation. * p

Figure Legend Snippet: Effects of azithromycin and amoxicillin + metronidazole (1:1 ratio) up to 5.0 mg/L on formation polymicrobial biofilms after 48 h of anaerobic incubation at 37°C in a 96-well plate model. Azithromycin and amoxicillin + metronidazole (1:1 ratio) at concentrations 0–100 mg/L were incubated with bacterial cultures. Data points represent the mean AU 620 value of a minimum of three biological replicates and the standard deviation. * p

Techniques Used: Incubation, Standard Deviation

Effect of amoxicillin + metronidazole up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Amoxicillin + metronidazole in a 1:1 ratio at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h at 37°C anaerobically. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

Figure Legend Snippet: Effect of amoxicillin + metronidazole up to 5.0 mg/L on the red complex mono- and polymicrobial biofilms in a 96-well plate model. Amoxicillin + metronidazole in a 1:1 ratio at concentrations 0–100 mg/L was incubated with bacterial cultures for 48 h at 37°C anaerobically. Data points represent the mean AU 620 value of a minimum of three biological replicates. Note the categorical scale.

Techniques Used: Incubation

4) Product Images from "Effect of azithromycin on a red complex polymicrobial biofilm"

Article Title: Effect of azithromycin on a red complex polymicrobial biofilm

Journal: Journal of Oral Microbiology

doi: 10.1080/20002297.2017.1339579

Techniques Used: Incubation

Techniques Used: Incubation, Staining

Techniques Used: Incubation, Standard Deviation

Techniques Used: Incubation

5) Product Images from "Intranasal Delivery of Cationic PLGA Nano/Microparticles- Loaded FMDV DNA Vaccine Encoding IL-6 Elicited Protective Immunity against FMDV Challenge"

Article Title: Intranasal Delivery of Cationic PLGA Nano/Microparticles- Loaded FMDV DNA Vaccine Encoding IL-6 Elicited Protective Immunity against FMDV Challenge

Journal: PLoS ONE

doi: 10.1371/journal.pone.0027605

Lymphocyte proliferation. Single lymphocyte suspensions were isolated from rats (n = 6) 7 days after the second immunization, plated in triplicate in a 96-well plate and stimulated in vitro for 48 h with inactivated FMDV, Con A (positive control) or with BSA. Means were compared by non-parametric ANOVA. Proliferation was analyzed using the MTT colorimetric assay and proliferation expressed as stimulated index. Significant results between pA-immunized animals and control groups are indicated by #.

Figure Legend Snippet: Lymphocyte proliferation. Single lymphocyte suspensions were isolated from rats (n = 6) 7 days after the second immunization, plated in triplicate in a 96-well plate and stimulated in vitro for 48 h with inactivated FMDV, Con A (positive control) or with BSA. Means were compared by non-parametric ANOVA. Proliferation was analyzed using the MTT colorimetric assay and proliferation expressed as stimulated index. Significant results between pA-immunized animals and control groups are indicated by #.

Techniques Used: Isolation, In Vitro, Positive Control, MTT Assay, Colorimetric Assay

6) Product Images from "Activated neutrophils exert myeloid-derived suppressor cell activity damaging T cells beyond repair"

Article Title: Activated neutrophils exert myeloid-derived suppressor cell activity damaging T cells beyond repair

Journal: Blood Advances

doi: 10.1182/bloodadvances.2019031609

Activated neutrophils suppress T-cell proliferation. Purified T cells (either CD4 + or CD8 + ) were cultured in the presence or absence of anti-CD3 antibody or anti-CD28 antibody with unstimulated or fMLF-activated neutrophils (unless otherwise indicated). Cells were harvested after 5 to 6 days and analyzed by flow cytometry for CFSE dilution. (A) Representative fluorescence-activated cell sorting (FACS) plots of CFSE dilution of CD4 + T cells. (B) Quantification of CD4 + (left) and CD8 + (right) T-cell proliferation (n = 17). (C) Titration of the cell ratio with 4000 (5:1 ratio), 20 000 (1:1), 40 000 (1:2), 60 000 (1:3), 100 000 (1:5), or 160 000 (1:8) neutrophils per well of a 96-well plate (n = 3-17). (D) Purified T cells were cultured in the presence or absence of IL-15 with unstimulated or fMLF-activated neutrophils (n = 5). (E) Purified T cells were cultured with anti-CD3 and anti-CD28 antibodies (red bars), and in the presence of neutrophils (blue bars) and/or indicated stimuli. Three to 19 donors were tested in duplicate per stimulus. Error bars indicate standard error of the mean (SEM); **** P

Figure Legend Snippet: Activated neutrophils suppress T-cell proliferation. Purified T cells (either CD4 + or CD8 + ) were cultured in the presence or absence of anti-CD3 antibody or anti-CD28 antibody with unstimulated or fMLF-activated neutrophils (unless otherwise indicated). Cells were harvested after 5 to 6 days and analyzed by flow cytometry for CFSE dilution. (A) Representative fluorescence-activated cell sorting (FACS) plots of CFSE dilution of CD4 + T cells. (B) Quantification of CD4 + (left) and CD8 + (right) T-cell proliferation (n = 17). (C) Titration of the cell ratio with 4000 (5:1 ratio), 20 000 (1:1), 40 000 (1:2), 60 000 (1:3), 100 000 (1:5), or 160 000 (1:8) neutrophils per well of a 96-well plate (n = 3-17). (D) Purified T cells were cultured in the presence or absence of IL-15 with unstimulated or fMLF-activated neutrophils (n = 5). (E) Purified T cells were cultured with anti-CD3 and anti-CD28 antibodies (red bars), and in the presence of neutrophils (blue bars) and/or indicated stimuli. Three to 19 donors were tested in duplicate per stimulus. Error bars indicate standard error of the mean (SEM); **** P

Techniques Used: Purification, Cell Culture, Flow Cytometry, Cytometry, Fluorescence, FACS, Titration

7) Product Images from "Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro"

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

Journal: bioRxiv

doi: 10.1101/2020.12.02.408112

Potency of selected HCV PI against SARS-CoV-2 was confirmed in Huh7.5 cells. Huh7.5 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir and simeprevir, as described in Materials and Methods. After 70-74 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of 7 replicates ± SEM and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay as described in Materials and Methods. Data points (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance of 12 nontreated controls. Sigmoidal concentration response curves were fitted and CC50 values were determined, as shown in Supplementary Figure 4. The blue stippled line represents the drug concentrations at which DMSO is expected to induce cytotoxicity with reduction of cell viability to

Figure Legend Snippet: Potency of selected HCV PI against SARS-CoV-2 was confirmed in Huh7.5 cells. Huh7.5 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir and simeprevir, as described in Materials and Methods. After 70-74 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of 7 replicates ± SEM and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay as described in Materials and Methods. Data points (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance of 12 nontreated controls. Sigmoidal concentration response curves were fitted and CC50 values were determined, as shown in Supplementary Figure 4. The blue stippled line represents the drug concentrations at which DMSO is expected to induce cytotoxicity with reduction of cell viability to

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Analysis of interactions of selected HCV PI with remdesivir. VeroE6 cells seeded in 96-well plates were infected the following day with SARS-CoV-2 followed by treatment with specified concentrations of the linear PI boceprevir (BOC) and narlaprevir (NAR), or the macrocyclic PI simeprevir (SIM), paritaprevir (PAR) and grazoprevir (GRA), or polymerase inhibitor remdesivir (REM), or a combination of these PI and remdesivir, as described in Materials and Methods. After 46-50 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Fractional effect (Fa) values were calculated by relating counts from infected and treated cultures to the mean count from at least 21 infected nontreated cultures and were entered into CompuSyn software. Datapoints are means of 6 to 7 replicates, and for each treatment experiment 6 to 10 datapoints were entered. For each inhibitor combination depicted per row, the following curves were fitted using Compusyn: (A) concentration-Fa curves plotting Fa values ranging from 0.01 to 0.99 against specified inhibitor concentrations. (B) Fa-CI curves plotting CI values ranging from 0 to 2 against Fa values ranging from 0.01 to 0.99. (C) Fa-Log 10 CI curves plotting logarithmic CI values ranging from 0.01 to 100 against Fa values ranging from 0.01 to 0.99. (B and C) Overall, CI values ≥1.1 suggest antagonism “A”, while CI values

Figure Legend Snippet: Analysis of interactions of selected HCV PI with remdesivir. VeroE6 cells seeded in 96-well plates were infected the following day with SARS-CoV-2 followed by treatment with specified concentrations of the linear PI boceprevir (BOC) and narlaprevir (NAR), or the macrocyclic PI simeprevir (SIM), paritaprevir (PAR) and grazoprevir (GRA), or polymerase inhibitor remdesivir (REM), or a combination of these PI and remdesivir, as described in Materials and Methods. After 46-50 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Fractional effect (Fa) values were calculated by relating counts from infected and treated cultures to the mean count from at least 21 infected nontreated cultures and were entered into CompuSyn software. Datapoints are means of 6 to 7 replicates, and for each treatment experiment 6 to 10 datapoints were entered. For each inhibitor combination depicted per row, the following curves were fitted using Compusyn: (A) concentration-Fa curves plotting Fa values ranging from 0.01 to 0.99 against specified inhibitor concentrations. (B) Fa-CI curves plotting CI values ranging from 0 to 2 against Fa values ranging from 0.01 to 0.99. (C) Fa-Log 10 CI curves plotting logarithmic CI values ranging from 0.01 to 100 against Fa values ranging from 0.01 to 0.99. (B and C) Overall, CI values ≥1.1 suggest antagonism “A”, while CI values

Techniques Used: Infection, Incubation, Immunostaining, Software, Concentration Assay

Potency of a panel of HCV PI and an HCV NS4A inhibitor against SARS-CoV-2 in VeroE6 cells. VeroE6 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir, narlaprevir, simeprevir, paritaprevir, grazoprevir, glecaprevir, voxilaprevir, vaniprevir, danoprevir, deldeprevir, asunaprevir and faldaprevir, as well as HCV NS4A inhibitor ACH-806, as described in Materials and Methods. After 46-50 hours of incubation, SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of counts from 7 replicate cultures ± standard errors of the means (SEM) and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay, as described in Materials and Methods. Datapoints (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance from 12 replicate nontreated control cultures. Sigmoidal concentration response curves were fitted and CC50 values were determined as shown in Supplementary Figure 3. The red / blue stippled line represents the drug concentrations at which DMSO is expected to induce antiviral effects with reduction of residual infectivity to

Figure Legend Snippet: Potency of a panel of HCV PI and an HCV NS4A inhibitor against SARS-CoV-2 in VeroE6 cells. VeroE6 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir, narlaprevir, simeprevir, paritaprevir, grazoprevir, glecaprevir, voxilaprevir, vaniprevir, danoprevir, deldeprevir, asunaprevir and faldaprevir, as well as HCV NS4A inhibitor ACH-806, as described in Materials and Methods. After 46-50 hours of incubation, SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of counts from 7 replicate cultures ± standard errors of the means (SEM) and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay, as described in Materials and Methods. Datapoints (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance from 12 replicate nontreated control cultures. Sigmoidal concentration response curves were fitted and CC50 values were determined as shown in Supplementary Figure 3. The red / blue stippled line represents the drug concentrations at which DMSO is expected to induce antiviral effects with reduction of residual infectivity to

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

8) Product Images from "Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity"

Article Title: Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity

Journal: Journal of Immunology Research

doi: 10.1155/2016/9540975

The luciferase IVT RNA-based assay efficiently assesses mAb-induced ADCC and CDC of tumour cell lines. ADCC assay using (a) KATO-III and (b) NUGC-4 cells. KATO-III and NUGC-4 cells endogenously expressing hCLDN18.2 were transfected with 7 μ g luc2 IVT RNA and seeded into 96-well plates independently. 4 h later, IMAB 362 at different concentrations and human PBMCs (E : T ratio = 40 : 1) from 6 different donors were added to the target cells and incubated for 24 h. ADCC was determined 40 and 45 min after addition of D-luciferin substrate to the KATO-III and NUGC-4 cells, respectively. (c) CDC assay. CHO-K1 cells stably expressing hCLDN18.2 were transfected with 7 μ g luc2 IVT RNA and seeded into 96-well plates. 24 h later, cells were incubated for 80 min with IMAB 362 diluted in human serum (final concentration of 20%) from 6 different healthy donors. CDC was determined 45 min after addition of D-luciferin substrate. Results are the mean ± SD ( n = 3).

Figure Legend Snippet: The luciferase IVT RNA-based assay efficiently assesses mAb-induced ADCC and CDC of tumour cell lines. ADCC assay using (a) KATO-III and (b) NUGC-4 cells. KATO-III and NUGC-4 cells endogenously expressing hCLDN18.2 were transfected with 7 μ g luc2 IVT RNA and seeded into 96-well plates independently. 4 h later, IMAB 362 at different concentrations and human PBMCs (E : T ratio = 40 : 1) from 6 different donors were added to the target cells and incubated for 24 h. ADCC was determined 40 and 45 min after addition of D-luciferin substrate to the KATO-III and NUGC-4 cells, respectively. (c) CDC assay. CHO-K1 cells stably expressing hCLDN18.2 were transfected with 7 μ g luc2 IVT RNA and seeded into 96-well plates. 24 h later, cells were incubated for 80 min with IMAB 362 diluted in human serum (final concentration of 20%) from 6 different healthy donors. CDC was determined 45 min after addition of D-luciferin substrate. Results are the mean ± SD ( n = 3).

Techniques Used: Luciferase, ADCC Assay, Expressing, Transfection, Incubation, CDC Assay, Stable Transfection, Concentration Assay

Electroporation of firefly luciferase IVT RNA into DCs and K562-A2 cells is nontoxic and leads to strong and long-lasting gene expression without affecting target cell phenotype. (a) Optimised luc2 reporter vector: composed of a gene-optimized synthetic firefly luciferase reporter gene cloned in front of two human β -globin 3′ untranslated regions (UTRs) fused head to tail and an unmasked free poly(A) tail of 120 bp. (b) Kinetics of luc2 expression in K562-A2 cells ( n = 1), human iDCs ( n = 3), and mDCs ( n = 3). Cells transfected with 8 pmol of luc2 -encoding IVT RNA were harvested at different time points to measure luminescence from 1 × 10 4 cells (Bright-Glo Luciferase Assay Kit for 96-well plates (Promega)). Results are the mean ± SD luminescence. Percent luminescence is relative to the highest luminescence signal obtained in each experiment. (c) Viability, reporter gene expression of iDCs and mDCs after eGFP and luc2 electroporation and phenotype after electroporation are depicted in descending order, respectively. iDCs (left panel) and mDCs (right panel) of 2 different donors were transfected with 10 μ g eGFP- or luc2 -encoding IVT RNA. Negative controls: cells electroporated without RNA (mock) and unelectroporated (no e'p) cells. Cells were harvested at different time points. Viability and HLA-DR, CD83, and eGFP expression levels were determined by flow cytometry. Luciferase activity of 1 × 10 4 viable cells was measured by luminescence in triplicate.

Figure Legend Snippet: Electroporation of firefly luciferase IVT RNA into DCs and K562-A2 cells is nontoxic and leads to strong and long-lasting gene expression without affecting target cell phenotype. (a) Optimised luc2 reporter vector: composed of a gene-optimized synthetic firefly luciferase reporter gene cloned in front of two human β -globin 3′ untranslated regions (UTRs) fused head to tail and an unmasked free poly(A) tail of 120 bp. (b) Kinetics of luc2 expression in K562-A2 cells ( n = 1), human iDCs ( n = 3), and mDCs ( n = 3). Cells transfected with 8 pmol of luc2 -encoding IVT RNA were harvested at different time points to measure luminescence from 1 × 10 4 cells (Bright-Glo Luciferase Assay Kit for 96-well plates (Promega)). Results are the mean ± SD luminescence. Percent luminescence is relative to the highest luminescence signal obtained in each experiment. (c) Viability, reporter gene expression of iDCs and mDCs after eGFP and luc2 electroporation and phenotype after electroporation are depicted in descending order, respectively. iDCs (left panel) and mDCs (right panel) of 2 different donors were transfected with 10 μ g eGFP- or luc2 -encoding IVT RNA. Negative controls: cells electroporated without RNA (mock) and unelectroporated (no e'p) cells. Cells were harvested at different time points. Viability and HLA-DR, CD83, and eGFP expression levels were determined by flow cytometry. Luciferase activity of 1 × 10 4 viable cells was measured by luminescence in triplicate.

Techniques Used: Electroporation, Luciferase, Expressing, Plasmid Preparation, Clone Assay, Transfection, Flow Cytometry, Cytometry, Activity Assay

9) Product Images from "Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro"

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

Journal: bioRxiv

doi: 10.1101/2020.12.02.408112

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Techniques Used: Infection, Incubation, Immunostaining, Software, Concentration Assay

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

10) Product Images from "Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro"

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

Journal: bioRxiv

doi: 10.1101/2020.12.02.408112

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Techniques Used: Infection, Incubation, Immunostaining, Software, Concentration Assay

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

11) Product Images from "Serum Amyloid P Component Bound to Gram-Negative Bacteria Prevents Lipopolysaccharide-Mediated Classical Pathway Complement Activation"

Article Title: Serum Amyloid P Component Bound to Gram-Negative Bacteria Prevents Lipopolysaccharide-Mediated Classical Pathway Complement Activation

Journal: Infection and Immunity

doi:

SAP inhibits the lysis of serovar Copenhagen Re by serum. Log phase-grown serovar Copenhagen Re was incubated overnight in HBSS–0.2% albumin–10% Mueller-Hinton broth in different concentrations of SAP − serum in the absence or presence of SAP (20 μg/ml) at 37°C in 96-well flat-bottom plates. The increase in optical density at 665 nm (OD 665 ) was used to determine bacterial growth. Data are means for three separate experiments ± SEM.

Figure Legend Snippet: SAP inhibits the lysis of serovar Copenhagen Re by serum. Log phase-grown serovar Copenhagen Re was incubated overnight in HBSS–0.2% albumin–10% Mueller-Hinton broth in different concentrations of SAP − serum in the absence or presence of SAP (20 μg/ml) at 37°C in 96-well flat-bottom plates. The increase in optical density at 665 nm (OD 665 ) was used to determine bacterial growth. Data are means for three separate experiments ± SEM.

Techniques Used: Lysis, Incubation

12) Product Images from "Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro"

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

Journal: bioRxiv

doi: 10.1101/2020.12.02.408112

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Techniques Used: Infection, Incubation, Immunostaining, Software, Concentration Assay

Techniques Used: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Transfection:

Article Title: Intranasal Delivery of Cationic PLGA Nano/Microparticles- Loaded FMDV DNA Vaccine Encoding IL-6 Elicited Protective Immunity against FMDV Challenge
Article Snippet: .. Lysed cells normalized to the total protein content were added to 96-well flat-bottom plates and detection of IL-6 by ELISA carried out as described by the bovine IL-6 kit instruction (Thermo Fisher) just expressing the IL-6 titer from transfected cells in optical density (OD). .. Preparation of plasmid DNA (pDNA)-adsorbed CHT-PLGA Nanoparticles were prepared using the emulsion-diffusion-evaporation technique .

Binding Assay:

Article Title: Role for Bcl-xL as an inhibitor of cytosolic cytochrome C accumulation in DNA damage-induced apoptosis
Article Snippet: .. Cytochrome C was adsorbed onto the plastic surface of a 96-well ELISA plate, followed by addition of blocking buffer (Superblock, Pierce) to reduce nonspecific binding. ..

Cell Culture:

Article Title: Triptolide, a constituent of immunosuppressive Chinese herbal medicine, is a potent suppressor of dendritic-cell maturation and trafficking
Article Snippet: .. Purified CD4+ T cells (105 /well) were cultured in 96-well flat-bottom plates with CD3/CD28 T-cell expansion Dynabeads (5 × 103 /well; Dynal ASA, Oslo, Norway). ..

Purification:

Enzyme-linked Immunosorbent Assay:

Incubation:

Article Title: Identification and characterization of noncalcemic, tissue-selective, nonsecosteroidal vitamin D receptor modulators
Article Snippet: .. KerTr cells (human transformed skin keratinocyte, obtained from ATCC) were plated in 96-well flat-bottom plates (3000 cells per well) in 100 μl keratinocyte serum-free medium supplemented with bovine pituitary extract in the absence of EGF (Invitrogen Corp.) and incubated at 37°C for 2 days. .. The cells were treated with various concentrations of VDR ligands in triplicate, dissolved in 100 μl keratinocyte serum-free medium supplemented with bovine pituitary extract in the absence of EGF, and incubated at 37°C for 72 hours.

other:

Article Title: Blocking interaction between SHP2 and PD‐1 denotes a novel opportunity for developing PD‐1 inhibitors
Article Snippet: Cells were pretreated with DMSO or MB or 25 μg/ml pembrolizumab or 20 μg/ml nivolumab for 1 h and then seeded in a 96‐well plate precoated with 10 μg/ml of aCD3/aCD28, with media supplemented with 10 μg/ml of hPD‐L1 protein.

Blocking Assay:

Expressing:

Transformation Assay:

Recombinant:

Article Title: Generation and characterization of monoclonal antibodies against the N-terminus of alpha-2-antiplasmin
Article Snippet: .. 96-well microtiter plates (Nunc A/S, Roskilde, Denmark) were coated with 2 μg/ml (200 μl per well in PBS containing 137 mM NaCl, 2.7 mM KCL, 6.5 mM Na2 HPO4 and 1 mM KH2 PO4 (pH 7.4)) of recombinant Met(R6)-α2AP, Met(W6)-α2AP or Asn-α2AP for 24 hours at 4°C. .. Non-specific sites were blocked with 200 μl 1% bovine serum albumin (Sigma, Steinheim, Germany) in PBS and incubated for two hours at RT.

Plasmid Preparation:

Article Title: Inhibition of Pyrimidine Biosynthesis Pathway Suppresses Viral Growth through Innate Immunity
Article Snippet: .. For one 96-well culture plate, 17 µg of plasmid were diluted in 500 µl of DMEM (Gibco-Invitrogen). .. In parallel, 53 µg of poly(ethyleneimine) from Sigma-Aldrich (PEI) were diluted in 500 µl of DMEM (Gibco-BRL).

96 well flat bottom plates (Thermo Fisher)

Structured Review

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Transfection:

Binding Assay:

Cell Culture:

Purification:

Enzyme-linked Immunosorbent Assay:

Incubation:

other:

Blocking Assay:

Expressing:

Transformation Assay:

Recombinant:

Plasmid Preparation:

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