HIV pseudovirus (PSV) was produced in HEK 293 T cells (ATCC, Manassas, VA, CRL-3216) and used in TZM-bl neutralization assays as previously described (Goo et al., 2012 (link)). Briefly, monoclonal antibodies were incubated with PSV at 37°C for 1 hr and TZM-bl cells (obtained through the NIH AIDS Reagent Program, Germantown, MD, catalog #8129 from Dr. John C. Kappes, and Dr. Xiaoyun Wu) were added at 10,000 cells/well and the assay was harvested after 48 hr at 37°C by adding Gal-Screen substrate (Thermo Fisher Scientific, catalog #T1028) as previously described (Simonich et al., 2019 (link)). IC50 values represent the half maximal inhibitory concentration in ug mL−1 at which 50% of the virus was neutralized. All neutralization data are the average of at least two independent experiments, each performed in technical duplicate (see Figure 2—source data 1 ; Figure 4—source data 1 ; Figure 5—source data 1 ; Figure 6—source data 1 ; Figure 7—source data 1 ; Figure 8—source data 1 ). IC50 values that deviated from the technical replicates by >10-fold were excluded as outliers from the average IC50 calculation.
Gal screen substrate
Manufactured by Thermo Fisher Scientific
Gal-Screen substrate is a chromogenic substrate used for the detection and quantification of beta-galactosidase activity in various biological applications. It provides a colorimetric readout upon enzymatic cleavage, allowing for the visualization and measurement of beta-galactosidase levels.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using gal screen substrate
1
Pseudovirus Neutralization Assay Protocol
2
HIV Pseudovirus Neutralization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HIV pseudovirus (PSV) was produced in HEK 293-T cells (ATCC®, Manassas, VA, CRL-3216™) and used in TZM-bl neutralization assays as previously described (Goo et al., 2012) (link).
Briefly, monoclonal antibodies were incubated with PSV at 37˚C for one hour and TZM-bl cells (obtained through the NIH AIDS Reagent Program, Germantown, MD, catalog #8129 from Dr.
John C. Kappes, and Dr. Xiaoyun Wu) were added at 10,000 cells/well and the assay was harvested after 48 hours at 37˚C by adding Gal-Screen substrate (Thermo Fisher Scientific, catalog #T1028) as previously described (Simonich et al., 2019) (link). IC50 values represent the half maximal inhibitory concentration in ug mL -1 at which 50% of the virus was neutralized. All neutralization data are the average of at least two independent experiments, each performed in technical duplicate (see Figures 2,4-7 -Neut Source Data 1). IC50 values that deviated from the technical replicates by >10-fold were excluded as outliers from the average IC50 calculation.
Briefly, monoclonal antibodies were incubated with PSV at 37˚C for one hour and TZM-bl cells (obtained through the NIH AIDS Reagent Program, Germantown, MD, catalog #8129 from Dr.
John C. Kappes, and Dr. Xiaoyun Wu) were added at 10,000 cells/well and the assay was harvested after 48 hours at 37˚C by adding Gal-Screen substrate (Thermo Fisher Scientific, catalog #T1028) as previously described (Simonich et al., 2019) (link). IC50 values represent the half maximal inhibitory concentration in ug mL -1 at which 50% of the virus was neutralized. All neutralization data are the average of at least two independent experiments, each performed in technical duplicate (see Figures 2,4-7 -Neut Source Data 1). IC50 values that deviated from the technical replicates by >10-fold were excluded as outliers from the average IC50 calculation.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!