Lanthascreen Cellular Pathway Assays

LanthaScreen™ STAT5 TF-1 Cell Line Thermo Scientific™

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. In hematopoeitic cells, the JAK2⁄STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. In this pathway, binding of these cytokines to their respective cell surface receptors results in the activation of JAK2, which in turn phospho-activates STAT5 proteins at specific tyrosine residues (Tyr-694⁄699). LanthaScreenTM-STAT5 TF-1 is a human hematopoeitic cell line which constitutively expresses GFP-STAT5 fusion proteins. The JAK2⁄STAT5 signaling pathway is known to be functionally intact in this cell line, therefore the GFP-STAT5 fusion protein serves as a substrate for the inducible phosphorylation by JAK2. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-STAT5 is detected in cell lysates using a LanthaScreenTM Terbium-anti-mouse and anti-phospho STAT5 [pTyr694⁄699] antibody pair, in a time-resolved FRET (TR-FRET) readout. GFP-STAT5 Lentivirus was transduced into TF-1 cells followed by selection with Blasticidine. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 fusion protein. Using a lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z' under optimized conditions using GM-CSF as an agonist for JAK2 -mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis⁄equilibration time.

LanthaScreen™ Tb-anti-LRRK2 [pSer935] Antibody Thermo Scientific™

The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody enables researchers to interrogate phosphorylation at serine 935 of the Leucine-Rich Repeat Kinase-2 (LRRK2) protein in a cellular, high-throughput screening (HTS)-compatible, and TR-FRET format.

This antibody has been specifically developed and optimized for use in our LanthaScreen® TR-FRET LRRK2 Cellular Activity Assay Kits (available for both wildtype and disease relevant mutant protein), which utilize the BacMam gene delivery system. The antibody, as well as other components of these assay kits, are also available separately. This assay format provides a convenient tool for the expression of LRRK2-GFP fusion protein in the cell background of your choice. The resulting cellular assay can be used to identify LRRK2 and relevant mutant inhibitor.

This product includes:

• LanthaScreen® Tb-anti-LRRK2 [pSer935] Antibody, 1 mg (enough to perform 275 384-well plate assays)

Additional products needed for cell-based assay:

BacMam LRRK2-GFP Wild Type Reagent, 15 mL or BacMam LRRK2-GFP G2019S Reagent, 15 mL
LanthaScreen® 6x Lysis Buffer, 50 mL
Instrument Control Terbium TR-FRET Kit

Benefits of this assay include:

More Physiologically Relevant: cell-based LRRK2 kinase activity and choice of cell background
More Convenient: compatible with High Throughput Screening (HTS); LanthaScreen® technology reduces interference and noise
Disease Relevant: screen compound libraries for Parkinson's disease-relevant molecules

More Physiologically Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody allows the investigation of phosphorylation at serine 935 on LRRK2 protein kinase. When paired with LRRK2 BacMam reagents, you have the freedom to choose the cellular background for your assay, including primary cells. This enables screening for potential inhibitors of LRRK2 proteins in their natural complexes in a physiologically relevant cell type.

More Convenient
Assays are run in a fully homogenous, addition-only format without any of the washing, lysate transfer, or separation procedures required for traditional methods such as western blotting and ELISA, making this assay ideal for HTS applications. The LanthaScreen® technology provides all of the advantages of TR-FRET ratiometric detection, including reduced data noise, less interference from fluorescent compounds, and high sensitivity, which results in the use of fewer cells than traditional Western or ELISA methods.

Disease Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody can be used by Parkinson's and other disease researchers to perform HTS, cell-based assays to generate inhibition curves (IC50) for their compound libraries. This will aid drug discovery efforts by reducing the time to results from 4 hrs to typically 1.5 hrs compared to alternative ELISAs.

For Research Use Only. Not intended for animal or human diagnostic or therapeutic use.

Related Links:
LRRK2 tools for advancing Parkinson's disease research
Kinase protein portfolio
LanthaScreen® Eu Kinase Binding Assay
LanthaScreen® Activity Assay
Learn More About BacMam Technology

LanthaScreen™ Akt HEK 293E Cell Line Thermo Scientific™

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. The PI3K⁄AKT pathway mediates signals for cell growth, cell survival, transcription, translation, and glucose uptake. Because of the complexity of this signaling cascade, especially as applied to the regulation of the mammalian target of rapamycin (mTOR), cell-based methods are critical for proper identification of small-molecule mediators of this pathway. The significance of mTOR as a kinase has been underscored recently by the identification of two distinct multimeric complexes inside the cell: mTORC1 (includes raptor and is rapamycin sensitive) and mTORC2 (includes rictor and is insensitive to acute rapamycin exposure). mTORC2 has been shown to phosphorylate and AKT at residue Ser473 for complete activation of this prosurvival kinase. LanthaScreenTM AKT HEK293E is a human cell line which constitutively expresses GFP-AKT fusion proteins. This kinase target was introduced using lipid transfection and these cells are a clonal population isolated by FACS, using GFP fluorescence as a sorting marker and Blasticidin to maintain cells under selection. Using this cell line, a homogenous immunoassay was developed with a time-resolved FRET (TR-FRET) readout in which the insulin-induced phosphorylation of Ser473 on GFP-AKT is detected in cell lysates using a terbium-labeled phosphor-specific antibody. This cell line has been validated with different stimuli⁄inhibitors and shows correct EC50 ⁄ IC50 values. Moreover, this assay has been optimized for performance under variable experimental conditions (including cell plating density, agonist stimulation time, DMSO tolerance and assay development time) and displays excellent statistical data (Z' > 0.6) and good signal-to-background.

LanthaScreen™ PRAS40 293E Cell Line Thermo Scientific™

The PI3K/AKT pathway is activated by various stimuli (including insulin and IGF-1) and mediates signals for cell growth, cell survival, translation, and inhibition of apoptosis. PRAS40 (Proline-rich AKT substrate, 40 kDa) is a cytosolic protein that is phosphorylated at position Thr246 by AKT directly upon stimulation of the pathway. PRAS40 has been shown to bind to the mTORC1 complex and inhibit downstream signaling (i.e., phosphorylation of 4E-BP1 or p70S6K). LanthaScreen® PRAS40 HEK293E is a human cell line which constitutively expresses GFP-PRAS40 fusion proteins. The kinase substrate
was introduced using lentiviral transduction followed by selection with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. The PI3K/AKT pathway is known to be active in the highly insulin-responsive HEK293E cell background. Using this cell line, a homogenous ('addition only') immunoassay has been developed in a 384-well format. The phosphorylation of PRAS40 at Thr246 is detected in cell lysates using a terbium-labeled phosphospecific antibody in a time-resolved FRET (TR-FRET) readout. The performance of this assay has been tested using numerous experimental variables, including cell plating density, stimulation time, DMSO tolerance, and antibody incubation time. Under optimized conditions, the assay has been further validated and shows correct EC50 and IC50 values, with insulin as the primary agonist and PI-103 as the known inhibitor. Overall, the assay displays excellent statistical data (Z'-factor >0.6), high signal-to-background (response ratio), and is a robust cell-based readout of AKT signaling.

LanthaScreen™ PDCD4 HEK 293E Cell Line Thermo Scientific™

The PI3K⁄AKT⁄mTOR pathway mediates signals for cell growth and survival, transcription and translation, and regulated cell death. The significance of mTOR has been underscored recently by the identification of two distinct cellular complexes: mTORC1 (includes raptor and is rapamycin sensitive) and mTORC2 (includes rictor and is insensitive to rapamycin). Activation of the pathway by insulin (or other mitogens) leads to the phosphorylation of programmed cell death protein 4 (PDCD4) via mTORC1 (indirectly through the ribosomal protein p70 S6 kinase). LanthaScreen® PDCD4 HEK293E is a human cell line which constitutively expresses GFP-PDCD4 fusion proteins. This kinase target was introduced using lipid transfection followed by selection with Blasticidin. These cells are a clonal population isolated by FACS using GFP fluorescence as a sorting marker. A homogenous immunoassay ('addition only', no wash steps) was developed in 384-well format using a TR FRET-based detection method to monitor the phosphorylation of Ser457 on GFP-PDCD4 using a terbium-labeled phosphospecific antibody. The assay has been optimized for performance under numerous experimental conditions (e.g., cell plating density, DMSO tolerance, insulin stimulation time, etc.) and displays excellent statistical data (Z' > 0.6). Moreover, this cell line has been validated with different stimuli and known inhibitors (shows correct EC50 and IC50 values) and serves as robust cellular readout for mTORC1 signaling.

LanthaScreen™ ERK2 U2OS Cell Line Thermo Scientific™

The Map Kinase (MapK) signal transduction cascade is activated by growth factors such as EGF, PDGF, and HGF. Binding of these factors to their respective cell surface receptors results in the initiation of receptor tyrosine kinase activity, which leads to the sequential phospho-activation of downstream kinases such as Ras, Raf, MEK, and Erk1⁄2. Activated MEK phosphorylates Erk2 (Mapk1) proteins at a specific Thr-Tyr motif (Thr⁄Tyr 185⁄187). LanthaScreenTM Erk2 U2OS is a human cell line which constitutively expresses GFP-Erk2. The MapK signaling pathway is functionally intact in this cell line, therefore the GFP-Erk2 fusion protein serves as a substrate for the growth-factor-inducible phosphorylation by MEK. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-Erk2 is detected in cell lysates using a terbium-labeled anti-pTpY-185⁄187-Erk2 antibody, in a time-resolved FRET (TR-FRET) readout. GFP-Erk2 lentivirus was transduced into U2OS cells, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. Using the lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z’ under optimized conditions using EGF as a ligand for GFP-Erk2 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and cell lysis⁄equilibration time. Additional information using alternate stimuli and small molecule inhibitor is also provided.

LanthaScreen™ STAT3 GripTite™ Cell Line Thermo Scientific™

The JAK/STAT3 signaling pathway is known to be activated by cytokines such as IL-6. In this pathway, binding of IL-6 to its cell-surface receptors results in the activation of JAKs, which in turn phosphoactivate STAT3 proteins at a specific tyrosine residue (Y705). LanthaScreen® STAT3 GripTite™ is a human cell line that constitutively expresses a GFP-STAT3 fusion protein. The JAK/STAT signaling pathway is known to be functionally intact in this cell line; therefore, the GFP-STAT3 fusion protein serves as a substrate for IL-6-inducible phosphorylation. Using this cell line, a lytic immunoassay has been developed in which the phosphorylation state of GFP-STAT3 is detected in cell lysates using a terbium-labeled anti-pY705-STAT3 antibody in a time-resolved FRET (TR-FRET) readout.

The GFP-STAT3 DNA expression construct was transfected into the GripTite™ HEK 293 cell line using Lipofectamine™ 2000 transfection reagent, and the transfected cells were selected with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. The assay utilizing this cell line is validated for EC50 and Z’-factors under optimized conditions using IL-6 as a ligand for JAK-mediated GFP-STAT3 phosphorylation. This assay has also been tested under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance, and assay equilibration time. An alternate "mix-and-read" assay format is also described.

LanthaScreen™ STAT1 U2OS Cell Line Thermo Scientific™

The JAK/STAT1 signaling pathway is known to be activated by type I/II interferons such as interferon-gamma and interferon-alpha. In this pathway, binding of these cytokines to their respective cell-surface receptors results in the activation of JAKs, which in turn phosphoactivate STAT1 proteins at a specific tyrosine residue (Y701). LanthaScreen® STAT1 U2OS is a human cell line that constitutively expresses a GFPSTAT1 fusion protein. The JAK/STAT1 signaling pathway is known to be functionally intact in this cell line; therefore, the GFP-STAT1 fusion protein serves as a substrate for the IFN-gamma-inducible phosphorylation by JAKs. Using this cell line, a homogeneous immunoassay has been developed in which the phosphorylation state of GFP-STAT1 is detected in cell lysates using a terbium-labeled anti-pY701-STAT1 antibody in a time-resolved FRET (TR-FRET) readout. The GFP-STAT1 DNA expression construct was transfected into U2OS cells using Lipofectamine™ LTX Reagent, followed by selection with blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as a sorting marker. Using the lytic TR-FRET immunoassay, this cell line is validated for EC50 and Zfi-factors under optimized conditions using IFN-gamma as a ligand for JAK-mediated GFP-STAT1 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance, and assay development time. Additional information using alternate stimuli and alternate assay protocols is available.

LanthaScreen™ ATF2 (19-106) A549 Cell Line Thermo Scientific™

The LanthaScreen®ATF2 (19-106) A549 cell line contains a fusion protein consisting of GFP and a fragment encoding for aa 19-106 of ATF2 under the control of a CMV promotor. A549 is a lung carcinoma cell line which shows inducible activation of JNK by number of different ligands, such as TNF and EGF, which leads to the transient phosphorylation of GFP-ATF2 (19-106). This LanthaScreen®cell line therefore allows the analysis of JNK activity using GFP-ATF2 phosphorylation as readout. The GFP-ATF2 (19-106) construct was transfected into A549 cells using Lipofectamine™2000, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. This cell line is validated for EC50 and Z' under optimized conditions using TNF as ligand for JNK mediated GFP-ATF2 (19-106) phosphorylation. This cell line has been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and assay development time. Additional information using alternate stimuli is also available. Academic and non-profit customers, please inquire for special pricing.

LanthaScreen™ LRRK2 G2019S [pSer935] Cellular Assay Kit Thermo Scientific™

The LanthaScreen® LRRK2 G2019S [pSer935] Cellular Assay Kit combines the flexibility of the BacMam gene delivery system with the robustness and power of our LanthaScreen® TR-FRET technology. It allows researchers to interrogate phosphorylation at serine 935 on a disease-relevant mutant (G2019S) Leucine-Rich Repeat Kinase-2 (LRRK2) protein in a variety of cell backgrounds to identify LRRK2 kinase inhibitors.

This assay format provides a convenient tool for the expression of G2019S LRRK2-GFP fusion protein in the cell background of your choice. The resulting cellular assay can be used to identify LRRK2 G2019S inhibitors, aiding in Parkinson's disease and other research focuses.

This kit provides enough material to evaluate the technology in a 384-well plate; larger sizes of all reagents are available separately.

The kit includes:

• LanthaScreen® Tb-anti-LRRK2 [pSer935] Antibody, 5 µg
• BacMam LRRK2-GFP G2019S Reagent, 5 × 1 mL
• LanthaScreen® 6x Lysis Buffer, 6 mL
• Instrument Control Terbium TR-FRET kit

A kit for interrogating wild-type LRRK2 protein is also available.

Benefits of this assay include:

More Physiologically Relevant: cell-based LRRK2 kinase activity and choice of cell background
More Convenient: compatible with High Throughput Screening (HTS); LanthaScreen® technology reduces interference and noise
Disease Relevant: screen compound libraries for Parkinson's disease-relevant molecules

More Physiologically Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody allows the investigation of phosphorylation at serine 935 on LRRK2 protein kinase. When paired with LRRK2 BacMam reagents, you have the freedom to choose the cellular background for your assay, including primary cells. This enables screening for potential inhibitors of LRRK2 proteins in their natural complexes in a physiologically relevant cell type.

More Convenient
Assays are run in a fully homogenous, addition-only format without any of the washing, lysate transfer, or separation procedures required for traditional methods such as western blotting and ELISA, making this assay ideal for HTS applications. The LanthaScreen® technology provides all of the advantages of TR-FRET ratiometric detection, including reduced data noise, less interference from fluorescent compounds, and high sensitivity, which results in the use of fewer cells than traditional Western or ELISA methods.

Disease Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody can be used by Parkinson's and other disease researchers to perform HTS, cell-based assays to generate inhibition curves (IC50) for their compound libraries. This will aid drug discovery efforts by reducing the time to results from 4 hrs to typically 1.5 hrs compared to alternative ELISAs.

For Research Use Only. Not intended for animal or human diagnostic or therapeutic use.

Related Links:
LRRK2 tools for advancing Parkinson's disease research
Kinase protein portfolio
LanthaScreen® Eu Kinase Binding Assay
LanthaScreen® Activity Assay
Learn More About BacMam Technology

LanthaScreen™ IkB alpha GripTite™ Cell Line Thermo Scientific™

The LanthaScreen®IkB alpha GripTite® cell line contains a fusion protein consisting of the cDNA encoding for GFP and IkB (GFP is fused to the N-terminus of IkB alpha) under the control of a CMV promoter stably transfected into GripTite® cells. GripTite® is a modified HEK293 cell line, which is a human kidney cell line which shows inducible activation of the NFkB signaling pathway by a number of different ligands, such as TNF and IL-1. Activation of the NFkB pathway involves the activation of IKK resulting in the phosphorylation of its target protein IkB alpha. This LanthaScreen cell line allows therefore the analysis of IKK activity using GFP-IkB alpha phosphorylation as readout. The GFP-IkB alpha construct was transfected into GripTite® cells using Lipofectamine™2000, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. This cell line is validated for EC50 and Z' under optimized conditions using TNF as ligand for IKK mediated GFP-IkB alpha phosphorylation or ubiquitination. This cell line has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and assay development time. Additional information using alternate stimuli is also available. Academic and non-profit customers, please inquire for special pricing.

LanthaScreen™ ERK2 A375 Cell Line Thermo Scientific™

The Map Kinase (MapK) signal transduction cascade is activated by growth factors such as EGF, PDGF, and HGF. Binding of these factors to their respective cell surface receptors results in the initiation of receptor tyrosine kinase activity, which leads to the sequential phospho-activation of downstream kinases such as Ras, Raf, MEK, and Erk1⁄2. Activated MEK phosphorylates Erk2 (Mapk1) proteins at a specific Thr-Tyr motif (Thr⁄Tyr 185⁄187). A number of constitutive active kinase mutants within the MAP kinase pathway have been implicated in oncogenesis. One of these mutants is BRAF(V600E), which is the most predominant oncogenic BRAF mutant. The human melanoma cell line A375 endogenously expresses BRAF(V600E), which leads to the constitutive activation of the MAP kinase pathway and phosphorylation of Erk2 in the absence of ligands. LanthaScreenTM Erk2 A375 constitutively expresses GFP-Erk2 under control of a CMV promotor. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-Erk2 is detected in cell lysates using a terbium-labeled anti-pTpY-185⁄187-Erk2 antibody, in a time-resolved FRET (TR-FRET) readout. This cell line can be used to evaluate compound activity against BRAF(V600E). GFP-Erk2 lentivirus was transduced into A375 cells, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. Using the lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z’ under optimized conditions using EGF as a ligand for GFP-Erk2 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and cell lysis⁄equilibration time. Additional information using alternate stimuli and small molecule inhibitor is also provided.

LanthaScreen™ STAT5 (JAK2 V617F) U2OS Cell Line Thermo Scientific™

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. The JAK2⁄STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. The recent discovery of an activating mutation in JAK2 (V617F) present in high percentage of myeloproliferative disease (MPD) patients suggests that this mutant JAK2 activity is a potential therapeutic target for certain forms of MPD. The assay described in this summary makes use of a cell line engineered for expression of the constitutively-active mutant kinase JAK2 V617F. By co-expressing GFP-STAT5 in this background, the phosphorylation state of STAT5 (specifically residue Tyr 694⁄699) can be modulated with JAK2 V617F inhibitors and analyzed in cell lysates using an anti-STAT5 [pTyr 694⁄699] and LanthaScreenTM terbium-anti-mouse antibody pair. GFP-STAT5 Lentivirus was transduced into U2OS cells followed by selection with Blasticidin. The selected pool was then transfected with a GST-JAK2 V617F construct using LipofectamineTM LTX, followed by selection with Geneticin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 and GST-JAK2 V617F. Using a lytic TR-FRET immuno-assay, this cell line is validated for IC50 and Z' under optimized conditions using JAK Inhibitor I (Pyridone 6) as a small molecule inhibitor for JAK2 V617F-mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis⁄equilibration time.

LanthaScreen™ LRRK2 [pSer935] Cellular Assay Kit Thermo Scientific™

The LanthaScreen® LRRK2 [pSer935] Cellular Assay Kit combines the flexibility of the BacMam gene delivery system with the robustness and power of our LanthaScreen® TR-FRET technology. It allows researchers to interrogate phosphorylation at serine 935 on a wild type Leucine-Rich Repeat Kinase-2 (LRRK2) protein in a variety of cell backgrounds to identify LRRK2 kinase inhibitors.

This assay format provides a convenient tool for the expression of LRRK2-GFP fusion protein in the cell background of of your choice. The resulting cellular assay can be used to identify LRRK2 and mutant inhibitorss, aiding in Parkinson's disease research and other research areas.

This kit provides enough material to evaluate the technology in a 384-well plate; larger sizes of all reagents are available separately.

The kit includes:

• LanthaScreen® Tb-anti-LRRK2 [pSer935] Antibody, 5 µg
• BacMam LRRK2-GFP Wild Type Reagent, 5 × 1 mL
• LanthaScreen® 6x Lysis Buffer, 6 mL
• Instrument Control Terbium TR-FRET Kit

A kit for interrogating a mutant LRRK2 protein is also available.

Benefits of this assay include:

More Physiologically Relevant: cell-based LRRK2 kinase activity and choice of cell background
More Convenient: compatible with High Throughput Screening (HTS); LanthaScreen® technology reduces interference and noise
Disease Relevant: screen compound libraries for Parkinson's disease-relevant molecules

More Physiologically Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody allows the investigation of phosphorylation at serine 935 on LRRK2 protein kinase. When paired with LRRK2 BacMam reagents, you have the freedom to choose the cellular background for your assay, including primary cells. This enables screening for potential inhibitors of LRRK2 proteins in their natural complexes in a physiologically relevant cell type.

More Convenient
Assays are run in a fully homogenous, addition-only format without any of the washing, lysate transfer, or separation procedures required for traditional methods such as western blotting and ELISA, making this assay ideal for HTS applications. The LanthaScreen® technology provides all of the advantages of TR-FRET ratiometric detection, including reduced data noise, less interference from fluorescent compounds, and high sensitivity, which results in the use of fewer cells than traditional Western or ELISA methods.

Disease Relevant
The LanthaScreen® Tb-Anti-LRRK2 [pSer935] Antibody can be used by Parkinson's and other disease researchers to perform HTS, cell-based assays to generate inhibition curves (IC50) for their compound libraries. This will aid drug discovery efforts by reducing the time to results from 4 hrs to typically 1.5 hrs compared to alternative ELISAs.

For Research Use Only. Not intended for animal or human diagnostic or therapeutic use.

Related Links:
LRRK2 tools for advancing Parkinson's disease research
Kinase protein portfolio
LanthaScreen® Eu Kinase Binding Assay
LanthaScreen® Activity Assay
Learn More About BacMam Technology

LanthaScreen™ c-Jun (1-79) HeLa Cell Line Thermo Scientific™

The LanthaScreen®c-Jun (1-79) HeLa cell line contains a fusion protein consisting of GFP and a fragment encoding for AA 1-79 of c-Jun under the control of a CMV promoter stably transfected into HeLa cell lines. HeLa is a cervical cancer cell line which shows inducible activation of JNK by a number of different ligands, such as TNF-a and EGF, which leads to the transient phosphorylation of GFP-c-Jun (1-79). This LanthaScreen®cell line therefore allows the analysis of JNK activity using GFP-c-Jun phosphorylation as readout. The GFP-c-Jun (1-79) construct was transfected into HeLa cells using Lipofectamine™2000, followed by selection with Blasticidin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker. This cell line is validated for EC50 and Z' under optimized conditions using TNF-a as ligand for JNK mediated GFP-c-Jun (1-79) phosphorylation. This cell lines has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and assay development time. Additional information using alternate stimuli is also available. Academic and non-profit customers, please inquire for special pricing.
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