HTRF Human Total cGAS Detection Kit HTRF®

Associated products

The HTRF Human Total cGAS detection kit is designed to monitor the expression level of cellular cGAS.

All inclusive kit
Low sample consumption
No-wash
High sensitivity

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The HTRF Human Total cGAS detection kit is designed to monitor the expression level of cellular cGAS.

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Overview

Following pathogen infection and the binding of dsDNA to the cytoplasmic sensor cGAS, STING protein is phosphorylated by TBK1, enabling its binding to IRF3 which leads to IFNs type 1 production. The STING pathway is then switched off by STING degradation, involving autophagy.

In immuno-oncology, activating the STING pathway has shown promising anti-tumor effects in pre-clinical models and thus represents a therapeutic strategy to treat human cancer.

Benefits

SPECIFICITY
PRECISION
LOW SAMPLE CONSUMPTION

Validation of Total cGAS detection in siRNA-treated THP-1 cells

THP-1 cells were plated at 100,000 cells per well under 90µl in a 96-well plates in complete culture medium. For Lipofectamine treated samples,10µl of Lipofectamine 1%f in culture medium were added. For siRNA treated cells, a 10µl mixture of Lipofectamine® RNAiMax/siRNA for cGas was added. Cells were then incubated for 48 h and 72 h at 37°C, 5% CO2.

After incubation, cells were lysed with 33.3µL of supplemented lysis buffer #1 at 4X for 30 minutes at RT under gentle shaking, and 16µL of lysate were transferred into a low volume white microplate before adding 2µL of the HTRF d2 detection reagent and2 µL HTRF Eu-K detection reagent. The HTRF signal was recorded after an ON incubation.

Validation of Total cGAS detection in siRNA-treated THP-1 cells

Validation of Total cGAS detection in siRNA-treated HeLa cells

HELA cells were plated at 50,000 cells per well under 90µl in a 96-well plates in complete culture medium. For Lipofectamine treated samples,10µl of Lipofectamine 1%f in culture medium were added. For siRNA treated cells, a 10µl mixture of Lipofectamine® RNAiMax/siRNA for cGas was added. Cells were then incubated for 24 h and 48 h at 37°C, 5% CO2.

After incubation, cells were lysed with 90µL of supplemented lysis buffer #1 at 1X for 30 minutes at RT under gentle shaking, and 16µL of lysate were transferred into a low volume white microplate before adding 2µL of the HTRF d2 detection reagent and 2µL HTRF Eu-K detection reagent. The HTRF signal was recorded after an ON incubation.

Validation of Total cGAS detection in siRNA-treated HeLa cells

HTRF cellular phospho-protein assays

Physiologically relevant results fo fast flowing research - Flyers

Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences

Insider Tips for successful sample treatment - Technical Notes

Optimize your HTRF cell signaling assays on tissues

HTRF and WB compatible guidelines - Technical Notes

Best practices for analyzing tumor xenografts with HTRF phospho assays

Protocol for tumor xenograft analysis with HTRF - Technical Notes

Key guidelines to successful cell signaling experiments

Mastering the art of cell signaling assays optimization - Guides

HTRF® phospho-protein platform facilitates the dissection of signaling pathways involved in insulin resistance and metabolic disorders

Multi-tissue cellular modeling and anlysis of insulin signaling - Posters

HTRF® cell signaling platform combined with iCell® Hepatocytes

A solution for phospho-protein analysis in metabolic disorders - Posters

HTRF phospho-assays reveal subtle drug-induced effects

Detailed protocol and direct comparison with WB - Posters

Implement HTRF phospho-protein assays at every step of the drug discovery process, from in vitro to in vivo models

A single technology for 2D cells, 3D cells, and xenograft models - Posters

Simplify complex pathway dissection by combining the power of HTRF® cellular phospho-assays and the flexibility of the CyBi®-FeliX liquid handling system

PI3K/AKT/mTor translational control pathway - Posters

Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells

Analysis of a large panel of diverse biological samples and cellular models - Posters

From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery

One technology across all samples - Application Notes

HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts

Tumor xenograft analysis: HTRF versus Western blot - Application Notes

HTRF cell-based phospho-protein data normalization

Valuable guidelines for efficiently analyzing and interpreting results - Application Notes

HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers

Increased flexibility of phospho-assays - Application Notes

HTRF Alpha-tubulin Housekeeping kit

Properly interpret your compound effect - Application Notes

Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling

Analyse of PI3K/AKT/mTor translational control pathway - Application Notes

How to run a cell based phospho HTRF assay

What to expect at the bench - Videos

Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening

In collaboration with Bayer - Scientific Presentations

Unleash the potential of your phosphorylation research with HTRF

A fun video introducing you to phosphorylation assays with HTRF - Videos

How to run a cell based phospho HTRF assay

3' video to set up your Phospho assay - Videos

Guidelines for Cell Culture and Lysis in Different Formats Prior to HTRF Detection

Seeding and lysing recommendations for a number of cell culture vessels. - Technical Notes

Methodological Aspects of Homogeneous Time-Resolved Fluorescence (HTRF)

Learn how to reduce time and sample consumption - Application Notes

Assessment of drug efficacy and toxicity by combining innovative technologies

Combination of AlphaLISA®, HTRF®, or AlphaLISA® SureFire® Ultra™ immunoassays with the ATPlite™ 1step cell viability assay - Application Notes

Product Insert ATG14 Total Kit / 64ATG14TPEG-64ATG14TPEH

64ATG14TPEG-64ATG14TPEH - Product Insert

Product Insert cGAS Total Kit / 64CGASTPEG-64CGASTPEH

64CGASTPEG-64CGASTPEH - Product Insert

Safety Data Sheet (DEU) cGAS Total Kit / 64CGASTPEG