Psicheck-2 Dual Glo Luciferase Reporter Assay Protocol

Description: The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly. This reagent lyses the cells and contains a substrate for firefly luciferase to produce firefly luciferase luminescence. Next, the Renilla Luciferase Reagent is added to the same well. It quenches the firefly luciferase luminescence and provides the substrate for renilla luciferase to produce renilla luciferase luminescence. The light production of both reactions can be conveniently measured on a luminometer. This assay system has several features:• Sensitive: highly sensitive detection of firefly luciferase activity and Renilla luciferase activity.• Stable: the luciferase signal output is stable for more than one hour, providing flexibility with regard to incubation time• High-throughput: simple homogeneous protocol minimizes handling steps to support high-throughput screening applications• Compatibility: works well with a variety of common media containing 0-10% serum and phenol red.

Description: The Dual Luciferase (Firefly-Renilla) Assay System is designed to be used for high-throughput, rapid quantitation of both Firefly and Renilla luciferases from a single sample in mammalian cell culture. The Firefly Luciferase Reagent is first added to the cells in medium directly. This reagent lyses the cells and contains a substrate for firefly luciferase to produce firefly luciferase luminescence. Next, the Renilla Luciferase Reagent is added to the same well. It quenches the firefly luciferase luminescence and provides the substrate for renilla luciferase to produce renilla luciferase luminescence. The light production of both reactions can be conveniently measured on a luminometer. This assay system has several features:• Sensitive: highly sensitive detection of firefly luciferase activity and Renilla luciferase activity.• Stable: the luciferase signal output is stable for more than one hour, providing flexibility with regard to incubation time• High-throughput: simple homogeneous protocol minimizes handling steps to support high-throughput screening applications• Compatibility: works well with a variety of common media containing 0-10% serum and phenol red.

Description: The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1; with D614G mutation) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 D614G variant in a Biosafety Level 2 facility.The Spike (D614G) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) has been validated for use with target cells Vero-E6 and ACE2-HEK293 (BPS Bioscience #79951). Spike VSV Delta G is preferred over lentiviral-based spike pseudoviruses for use in cells such as Vero-E6 parental cells.

Description: The Spike (BA.2 Variant) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) was produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BA.2 mutations; see below for details) as the envelope glycoprotein instead of VSV-G. The pseudovirions contain the firefly luciferase gene; therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BA.2 Variant) (SARS-CoV-2) Pseudotyped VSV Delta G (Luciferase Reporter) can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 BA.2 variant in a Biosafety Level 2 facility.

Description: The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) was produced by re-expression of VSV-G as the envelope glycoprotein using the VSV Delta G system in which VSV-G is deleted. The pseudovirions contain the firefly luciferase gene; therefore, the VSV-G mediated cell entry can be measured via luciferase activity. The VSV-G Pseudotyped VSV Delta G (Luciferase Reporter) can be used as a positive control of transduction for other VSV pseudotypes containing the envelope glycoproteins of heterologous viruses in a Biosafety Level 2 facility.

Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1 omicron variant has additional mutations (K444T and N460K) based on the BA.5 variant. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1 variant in a Biosafety Level 2 facility._x000D_

_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K

Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BQ.1.1 omicron variant has additional mutations (R346T, K444T and N460K) based on the BA.5 variant. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BQ.1.1 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BQ.1.1, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BQ.1.1 variant in a Biosafety Level 2 facility._x000D_

_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion_x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BQ.1.1 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BQ.1.1 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, K444T, L452R, N460K, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K

Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of October 2022, several new BA.5 sub-lineages (e.g. BQ.1, BQ.1.1, BF.7) have been designated._x000D_The spike protein of BF.7 omicron variant has additional mutation R346T based on the BA.5 variant. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the Omicron BF.7 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (BF.7, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron BF.7 variant in a Biosafety Level 2 facility._x000D_

_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figures 2 and 3 in Validation Data, the Spike Omicron BF.7 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in BF.7 Omicron Variant:_x000D_Del69-70, T19I, LPPA24-27S, G142D, V213G, G339D, R346T, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K

Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and human ACE2 may offer protection against the viral infection. Omicron Variant was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. As of May 2022, Omicron variants were divided into seven distinct sub-lineages: BA.1, BA.1.1, BA.2, BA.3, BA.2.12.1, BA.4, and BA.5. As of January 2023, additional new sub-lineages (e.g. BQ.1, BQ.1.1, BF.7, XBB.1, XBB.1.5) have been designated._x000D_The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 Spike (Genbank Accession #QHD43416.1 containing all the XBB.1.5 mutations; see below for details) as the envelope glycoprotein instead of the commonly used VSV-G. These pseudovirions contain the firefly luciferase gene driven by a CMV promoter (Figure 1), therefore, the spike-mediated cell entry can be measured via luciferase activity. The Spike (XBB.1.5, Omicron Variant) (SARS-CoV-2) pseudovirus can be used to measure the activity of a neutralizing antibody against SARS-CoV-2 Omicron XBB.1.5 variant in a Biosafety Level 2 facility._x000D_

_x000D_Figure 1. Schematic of the Luciferase Reporter in SARS-CoV-2 Spike Pseudovirion._x000D_As shown in Figure 2, the Spike Omicron XBB.1.5 pseudovirus has been validated for use with ACE2-HEK293 target cells (which overexpress ACE2; BPS Bioscience #79951)._x000D_Spike Mutations in XBB.1.5 Omicron Variant: T19I, LPP24-26del, A27S, V83A, G142D, Y144del, H146Q, Q183E, V213E, G252V, G339H, R346T, L368I, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, V445P, G446S, N460K, S477N, T478K, E484A, F486P, F490S, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K

Description: Minimum order quantity: 1 unit of 1000TST

Description: Minimum order quantity: 1 unit of 1000AS

Description: Minimum order quantity: 1 unit of 1000AS

Description: The RARα Reporter Cellular Assay Pack provides all the key reagents required to monitor the activity of retinoic acid receptor alpha (RARα). The pack contains the RARα Reporter (Luc)-HEK293 Cell Line, a luciferase reporter cell line that contains a firefly luciferase gene under the control of retinoic acid response elements stably integrated into HEK293 cells along with full length human RARα (GenBank Accession No. NM_000964). This cell line is functionally validated for the response to the stimulation of all-trans retinoic acid, and the expression of RARα is confirmed by Western blotting._x000D_Additionally, the pack includes cell culture medium (Thaw Medium 6) that has been optimized for use with HEK293 cells. Thaw Medium 6 includes 10% fetal bovine serum and 1% Pen/Strep. Finally, the pack provides the ONE-Step™ Luciferase Detection System. This reagent provides highly sensitive, stable detection of firefly (Photinus pyralis) luciferase activity. The ONE-Step luciferase reagent can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.

Description: The RARβ Reporter Cellular Assay Pack provides all the key reagents required to monitor the activity of retinoic acid receptor beta (RARβ). The pack contains the RARβ Reporter (Luc)-HEK293 Cell Line, a luciferase reporter cell line that contains a firefly luciferase gene under the control of retinoic acid response elements stably integrated into HEK293 cells along with full length human RARα (GenBank Accession No. P10826-2). This cell line is functionally validated for the response to the stimulation of all-trans retinoic acid, and the expression of RARβ is confirmed by Western blotting._x000D_Additionally, the pack includes cell culture medium (Thaw Medium 6) that has been optimized for use with HEK293 cells. Thaw Medium 6 includes 10% fetal bovine serum and 1% Pen/Strep. Finally, the pack provides the ONE-Step™ Luciferase Detection System. This reagent provides highly sensitive, stable detection of firefly (Photinus pyralis) luciferase activity. The ONE-Step luciferase reagent can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.

Description: The RARγ Reporter Cellular Assay Pack provides all the key reagents required to monitor the activity of retinoic acid receptor gamma (RARγ). The pack contains the RARγ Reporter (Luc)-HEK293 Cell Line, a luciferase reporter cell line that contains a firefly luciferase gene under the control of retinoic acid response elements stably integrated into HEK293 cells along with full length human RARα (GenBank Accession No. P13631-1). This cell line is functionally validated for the response to the stimulation of all-trans retinoic acid, and the expression of RARγ is confirmed by Western blotting._x000D_Additionally, the pack includes cell culture medium (Thaw Medium 6) that has been optimized for use with HEK293 cells. Thaw Medium 6 includes 10% fetal bovine serum and 1% Pen/Strep. Finally, the pack provides the ONE-Step™ Luciferase Detection System. This reagent provides highly sensitive, stable detection of firefly (Photinus pyralis) luciferase activity. The ONE-Step luciferase reagent can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.