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Intellicyt ique high throughput flow cytometer
PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an <t>iQue</t> flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.
Ique High Throughput Flow Cytometer, supplied by Intellicyt, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/high+throughput+flow+cytometer/cytometer+flow+high+throughput/pmc13178664-216-27-26
Average 86 stars, based on 1 article reviews
ique high throughput flow cytometer - by Bioz Stars, 2026-10
86/100 stars

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1) Product Images from "Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge"

Article Title: Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI191440

PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an iQue flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.
Figure Legend Snippet: PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an iQue flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.

Techniques Used: Binding Assay, Expressing, Flow Cytometry, Transfection, Software, Negative Control

PreGn binding antibodies were added to preGn-expressing cells at saturating concentrations (20 μg/mL) for 30 minutes. Alexa Fluor 647–conjugated mouse or human antibodies were then added to cells at 5 μg/mL without a prior wash step. Cells were washed and immediately processed on an iQue flow cytometer. The values shown are the percentage of binding that occurred during competition compared with noncompeted binding of the mAb. This value was normalized to 100%. The values are also indicated by the dotted lines; above 40% indicated no competition, between 40% and 20% indicated partial competition, and below 20% indicated competition. Values shown are from 2 independent experiments with 3 replicates. Data represent mean ± SD.
Figure Legend Snippet: PreGn binding antibodies were added to preGn-expressing cells at saturating concentrations (20 μg/mL) for 30 minutes. Alexa Fluor 647–conjugated mouse or human antibodies were then added to cells at 5 μg/mL without a prior wash step. Cells were washed and immediately processed on an iQue flow cytometer. The values shown are the percentage of binding that occurred during competition compared with noncompeted binding of the mAb. This value was normalized to 100%. The values are also indicated by the dotted lines; above 40% indicated no competition, between 40% and 20% indicated partial competition, and below 20% indicated competition. Values shown are from 2 independent experiments with 3 replicates. Data represent mean ± SD.

Techniques Used: Binding Assay, Expressing, Flow Cytometry

Related Articles

Binding Assay:

Article Title: Compositions and methods for inhibition of alphavirus infection
Article Snippet: .. Binding of Mxra8-Fc to each mutant expressed in HEK-293T cells was determined by immunofluorescence detected with a high-throughput flow cytometer (Intellicyt), as previously described36. ..

Article Title: Anti-Ebola virus mAb 3A6 with unprecedented potency protects highly viremic animals from fatal outcome and physically lifts its glycoprotein target from the virion membrane
Article Snippet: Protein variants were cell surface-expressed for 22 h . The indicated mAbs were incubated with the cells for 1 h before an Alexa Fluor 488-conjugated secondary antibody (Thermo Fischer Scientific) was added. .. Antibody binding was assessed by detection of cellular fluorescence with a high throughput flow cytometer (Intellicyt, Albuquerque, NM, USA). ..

Mutagenesis:

Article Title: Compositions and methods for inhibition of alphavirus infection
Article Snippet: .. Binding of Mxra8-Fc to each mutant expressed in HEK-293T cells was determined by immunofluorescence detected with a high-throughput flow cytometer (Intellicyt), as previously described36. ..

Immunofluorescence:

Article Title: Compositions and methods for inhibition of alphavirus infection
Article Snippet: .. Binding of Mxra8-Fc to each mutant expressed in HEK-293T cells was determined by immunofluorescence detected with a high-throughput flow cytometer (Intellicyt), as previously described36. ..

Flow Cytometry:

Article Title: Compositions and methods for inhibition of alphavirus infection
Article Snippet: .. Binding of Mxra8-Fc to each mutant expressed in HEK-293T cells was determined by immunofluorescence detected with a high-throughput flow cytometer (Intellicyt), as previously described36. ..

Article Title: Complete Combinatorial Mutational Enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 hour at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from 4 different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Cells were washed twice with HBS blocking buffer and cell fluorescence was measured using an IntelliCyt high throughput flow cytometer. .. Cells were gated to exclude non-single cells, FITC labeling was used to select RBD-expressing cells, and Alexa647 labeling was used to quantify Fc-ACE2-binding cells.

Article Title: Pan-ebola virus neutralizing human antibodies and methods of use therefor
Article Snippet: The detailed algorithms used to interpret shotgun mutagenesis data were described previously (Davidson and Doranz, 2014). .. After washing, cellular fluorescence was detected using the Intellicyt high throughput flow cytometer (Intellicyt). ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Next, viral transduction was measured as neonGreen fluorescence using an IntelliCyt high throughput flow cytometer. .. Uninfected controls and the lentiviral particles expressing the RBD of the L strain were used to set the gates.

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 h at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from four different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

Article Title: Anti-Ebola virus mAb 3A6 with unprecedented potency protects highly viremic animals from fatal outcome and physically lifts its glycoprotein target from the virion membrane
Article Snippet: Protein variants were cell surface-expressed for 22 h . The indicated mAbs were incubated with the cells for 1 h before an Alexa Fluor 488-conjugated secondary antibody (Thermo Fischer Scientific) was added. .. Antibody binding was assessed by detection of cellular fluorescence with a high throughput flow cytometer (Intellicyt, Albuquerque, NM, USA). ..

Fluorescence:

Article Title: Complete Combinatorial Mutational Enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 hour at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from 4 different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Cells were washed twice with HBS blocking buffer and cell fluorescence was measured using an IntelliCyt high throughput flow cytometer. .. Cells were gated to exclude non-single cells, FITC labeling was used to select RBD-expressing cells, and Alexa647 labeling was used to quantify Fc-ACE2-binding cells.

Article Title: Pan-ebola virus neutralizing human antibodies and methods of use therefor
Article Snippet: The detailed algorithms used to interpret shotgun mutagenesis data were described previously (Davidson and Doranz, 2014). .. After washing, cellular fluorescence was detected using the Intellicyt high throughput flow cytometer (Intellicyt). ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Next, viral transduction was measured as neonGreen fluorescence using an IntelliCyt high throughput flow cytometer. .. Uninfected controls and the lentiviral particles expressing the RBD of the L strain were used to set the gates.

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 h at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from four different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

Article Title: Anti-Ebola virus mAb 3A6 with unprecedented potency protects highly viremic animals from fatal outcome and physically lifts its glycoprotein target from the virion membrane
Article Snippet: Protein variants were cell surface-expressed for 22 h . The indicated mAbs were incubated with the cells for 1 h before an Alexa Fluor 488-conjugated secondary antibody (Thermo Fischer Scientific) was added. .. Antibody binding was assessed by detection of cellular fluorescence with a high throughput flow cytometer (Intellicyt, Albuquerque, NM, USA). ..

High Throughput Screening Assay:

Article Title: Complete Combinatorial Mutational Enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 hour at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from 4 different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Cells were washed twice with HBS blocking buffer and cell fluorescence was measured using an IntelliCyt high throughput flow cytometer. .. Cells were gated to exclude non-single cells, FITC labeling was used to select RBD-expressing cells, and Alexa647 labeling was used to quantify Fc-ACE2-binding cells.

Article Title: Pan-ebola virus neutralizing human antibodies and methods of use therefor
Article Snippet: The detailed algorithms used to interpret shotgun mutagenesis data were described previously (Davidson and Doranz, 2014). .. After washing, cellular fluorescence was detected using the Intellicyt high throughput flow cytometer (Intellicyt). ..

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Next, viral transduction was measured as neonGreen fluorescence using an IntelliCyt high throughput flow cytometer. .. Uninfected controls and the lentiviral particles expressing the RBD of the L strain were used to set the gates.

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: To estimate the IC50s of the neutralizing antibodies with pseudotyped lentivirus expressing the RBD of the L strain, 10-fold dilutions of the antibodies were incubated with L strain pseudotyped lentivirus for 1 h at 37 C and subsequently added to ACE2+ HEK293 cells. .. NeonGreen fluorescence was analyzed using an IntelliCyt high throughput flow cytometer and the data from four different experiments were used to estimate the IC50 values using the Graphpad Prism version 9.0. ..

other:

Article Title: Anti-Ebola virus mAb 3A6 with unprecedented potency protects highly viremic animals from fatal outcome and physically lifts its glycoprotein target from the virion membrane
Article Snippet: Antibody binding was assessed by detection of cellular uorescence with a high throughput ow cytometer (Intellicyt, Page 15/29 Albuquerque, NM, USA).

Blocking Assay:

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Cells were washed twice with HBS blocking buffer and cell fluorescence was measured using an IntelliCyt high throughput flow cytometer. .. Cells were gated to exclude non-single cells, FITC labeling was used to select RBD-expressing cells, and Alexa647 labeling was used to quantify Fc-ACE2-binding cells.

Transduction:

Article Title: Complete combinatorial mutational enumeration of a protein functional site enables sequence-landscape mapping and identifies highly-mutated variants that retain activity.
Article Snippet: .. Next, viral transduction was measured as neonGreen fluorescence using an IntelliCyt high throughput flow cytometer. .. Uninfected controls and the lentiviral particles expressing the RBD of the L strain were used to set the gates.

Sample Throughput:

Article Title: Anti-Ebola virus mAb 3A6 with unprecedented potency protects highly viremic animals from fatal outcome and physically lifts its glycoprotein target from the virion membrane
Article Snippet: Protein variants were cell surface-expressed for 22 h . The indicated mAbs were incubated with the cells for 1 h before an Alexa Fluor 488-conjugated secondary antibody (Thermo Fischer Scientific) was added. .. Antibody binding was assessed by detection of cellular fluorescence with a high throughput flow cytometer (Intellicyt, Albuquerque, NM, USA). ..



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PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an iQue flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.

Journal: The Journal of Clinical Investigation

Article Title: Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge

doi: 10.1172/JCI191440

Figure Lengend Snippet: PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an iQue flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.

Article Snippet: After 7 days, LCL supernatants were screened for the presence of CCHFV Gc-, Gn-, or GP38-recognizing antibodies using IbAr10200 strain M segment–transfected Expi293F cells on the Intellicyt iQue high-throughput flow cytometer as described below.

Techniques: Binding Assay, Expressing, Flow Cytometry, Transfection, Software, Negative Control

PreGn binding antibodies were added to preGn-expressing cells at saturating concentrations (20 μg/mL) for 30 minutes. Alexa Fluor 647–conjugated mouse or human antibodies were then added to cells at 5 μg/mL without a prior wash step. Cells were washed and immediately processed on an iQue flow cytometer. The values shown are the percentage of binding that occurred during competition compared with noncompeted binding of the mAb. This value was normalized to 100%. The values are also indicated by the dotted lines; above 40% indicated no competition, between 40% and 20% indicated partial competition, and below 20% indicated competition. Values shown are from 2 independent experiments with 3 replicates. Data represent mean ± SD.

Journal: The Journal of Clinical Investigation

Article Title: Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge

doi: 10.1172/JCI191440

Figure Lengend Snippet: PreGn binding antibodies were added to preGn-expressing cells at saturating concentrations (20 μg/mL) for 30 minutes. Alexa Fluor 647–conjugated mouse or human antibodies were then added to cells at 5 μg/mL without a prior wash step. Cells were washed and immediately processed on an iQue flow cytometer. The values shown are the percentage of binding that occurred during competition compared with noncompeted binding of the mAb. This value was normalized to 100%. The values are also indicated by the dotted lines; above 40% indicated no competition, between 40% and 20% indicated partial competition, and below 20% indicated competition. Values shown are from 2 independent experiments with 3 replicates. Data represent mean ± SD.

Article Snippet: After 7 days, LCL supernatants were screened for the presence of CCHFV Gc-, Gn-, or GP38-recognizing antibodies using IbAr10200 strain M segment–transfected Expi293F cells on the Intellicyt iQue high-throughput flow cytometer as described below.

Techniques: Binding Assay, Expressing, Flow Cytometry