This protocol enables the cells' viability and confluence to be monitored. The Dual-Luciferase ® Reporter Assay or DLR is widely used to study gene transcription and regulation. As shown in Figure 1, the interaction between Tag1-SOS1 and Tag2-KRAS-WT is detected by using anti-Tag1-Terbium (HTRF donor) and anti-Tag2-XL665 (HTRF acceptor). Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED. 96-well culture plateThe assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Alpha Technology: The VICTOR Nivo system. • HTRF®-Certified Reader. FOR HTRF CERTIFIED READER For. 2. It enables you to gain insights you couldn’t achieve with detection measurements alone. CLARIOstar is equipped with a specific optical device which enables the measurement of. 8 µL cells + 4 µL compounds + 4 µL supplemented ® lysis buffer (4X) + 4 µL HTRF pre-mixed antibodies. → Incubate 2h (if the detection is not sensitive enough, it is better to incubate 20h). The SpectraMax M5e Reader offers the additional benefit of being certified for Cisbio Bioassays’ HTRF® technology. 5 EXAMPLE OF MAP PLATE . Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READER. Make sure the setup for Eu Cryptate is used. • HTRF plate validated (QC with reference HTRF assays) • Low to high volumes (10 µL to 350 µL) • Can be used with all HTRF compatible readersHTRF combines standard FRET technology with time-resolved measurement of fluorescence, eliminating short-lived background fluorescence. Lysates are then transferred to the assay plate for the detection of total-CDK4 by HTRF. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. Read & analyse. The VICTOR ® Nivo ™ system packs all popular detection modes into the industry’s smallest multimode reader footprint. 5 µL. MUTANT HTT STANDARD WORKING SOLUTIONSLysis buffer 4X for two-plate assay protocol on suspension cells Determine the amount of lysis buffer needed for the experiment. Incubate overnight at RT. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. Assay plates were incubated at RT and read at different time points thereafter on an HTRF ® certified. It can also be further streamlined toFOR HTRF CERTIFIED READER. Lysates are then transferred to the assay plate for the detection of total-Rb by HTRF. Highly compatible plates for HTRF. Use simpler, faster protocols. They can be used in a no-transfer. Nephelometers are dedicated instruments that can only detect scattering and are only available as standalone plate readers. To perform the assay, use white plate only. • HTRF®-Certified Reader. A microplate reader that can detect two or more applications is considered a multi-mode plate reader. FOR HTRF CERTIFIED READER. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. FOR HTRF CERTIFIED READER For more. ONE-PLATE ASSAY PROTOCOL: Small volume detection plate. HTRF is compatible with a large range of available microplate readers. 25,000 human Jurkat cells were plated in 96 well plates. + 4 µL HTRF ® pre-mixed antibo dies 2. Thanks to HTRF signal stability, the assay plate may be used to check several instruments, or. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READERHTRF Ratio Anti-Tag Antibody Cryptate Inhibitor-Red Inhibitor Purified Tagged Kinase Figure 2: Principle of HTRF kinase competition binding assay (IC 50 - KI determination) HTRF Dasatinib-Red Part # 62KB02REDC & 62KB02REDE Amount: 1 nmol (62KB02REDC) & 20 nmol (62KB02REDE) Concentration: 25 µM in DMSO Form:. These plates can be read on all HTRF certified readers, thus making their implementation in your experiments really easy Provider Format Part Numbers Recommended volume Type; Cisbio: 96-w low volume: 6PL96005 – 66PL96025 – 66PL96100: 20-30 µL:FOR HTRF CERTIFIED READER. NanoQuant Plate™ allows rapid, straightforward measurement of 16 x 2 µl samples simultaneously without calibration. Lysates are then transferred to the assay plate for the detection of total-STING by HTRF. Specialized filters allow for IR fluorescence. For more information. It delivers even higher sensitivity for time-resolved fluorescence (TRF) applications, and the Enhanced Security software option includes. 8 µL cells + 4 µL compounds + 4 µL supplemented ® lysis buffer (4X) + 4 µL HTRF pre-mixed antibodies. FOR HTRF CERTIFIED READER For. The FLUOstar Omega is the ideal plate reader for life science applications. The PHERAstar FS is an advanced HTS plate reader designed for all HTRF assays and features the following: All microplate formats up to 3456-well. The FLUOstar ® Omega is a multi-mode microplate reader with three detection modes. 10 μL of GLP1 receptor cells were dispensed into wells of. Conclusion. 8 µL cells + 4 µL compounds + 4 µL supplemented ® lysis buffer (4X) + 4 µL HTRF pre-mixed antibodies. (HTRF) • Certified for detection of Cisbio’s HTRF assays • GPCR assays (cAMP cell based, membrane based). " The PHERAstar FSX instrument is a high quality, reliable, plate reader. 96-well culture plateFOR HTRF CERTIFIED READER For more information about HTRF. On the SpectraMax i3x reader, install the HTRF cartridge, and wavelengths will be selected automatically when HTRF is selected as the Optical Configuration. For more information about HTRF ®The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. Plates can be loaded manually or plates can be loaded using 20 or 50-plate free access magazines which fit in a stacker. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. 3. Incubate 4h at RT. Bench top solutions for cell based assays. Incubate overnight at RT. For more information about HTRF ®The LUMIstar® Omega is BMG LABTECH's powerful dedicated luminescence microplate reader, providing highly sensitive flash and glow assay readings. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. Typically the system can detect absorbance, luminescence, fluorescence, and even make more specialized fluorescence measurements like time-resolved fluorescence (TRF) and fluorescence polarization (FP). 2400 for 665 and 620 with white plate HTRF® Europium cryptate donor / Red acceptor readout. View Brochure. TR-FRET was measured on the SpectraMax i3x and iD5 readers (see Table 1 for instrument settings). Read & analyse. Depending on the cell lines used, it may be necessary to dilute the cell lysate to ensure samples are within the assay linear range. The EnVision XCite multimode plate reader provides exceptional sensitivity across all detection technologies, while keeping you flexible with monochromator option and full upgradeability. For more information about HTRF. Data Analysis HTRF®-certified, DLReady, and SpectraMax® DuoLuc™-certified. Lysates are then transferred to the assay plate for the detection of total cRaf by HTRF. Whatever the plate type, assay volume should match well volume as closely as possible so that focalization of the reader excitation beam gives maximum energy delivery (Fig. Microplate reader assays requiring infrared (IR) or red-shifted assays can be read with great sensitivity using the iD5 reader’s filter system. Cover the plate with a plate sealer. Make sure the setup for Eu3+ Cryptate is used. e. Thanks to HTRF signal stability, the assay plate can be used to check several instruments, or different reader. Preconfigured HTRF protocols in SoftMax ® Pro Software 7. Certified SpectraMax® M5e-HTRF Readers The SpectraMax M5 e reader has the same perfor mance specifications as the M5 but is certified for use with Cisbio Bioassays’ HTRF (Homogeneous Time-Resolved Fluorescence) technology. Highly compatible plates for HTRF. If not, our scientific team will help you choose the instrument that best suits your needs and budget. As shown in Figure 1, the interaction between Tag1-SOS1 and Tag2-KRAS-WT is detected by using anti-Tag1-Terbium (HTRF donor) and anti-Tag2-XL665 (HTRF acceptor). 8 µL cells + 4 µL compounds + 4 µL supplemented ® lysis buffer (4X) + 4 µL HTRF pre-mixed antibodies. White 384-well low volume plate Make sure to use the set-up for Eu 3+ Cryptate. FOR HTRF CERTIFIED READER . The plate was covered both incubated for one hour at room temperature. Plate Reader Requirement. The EnVision XCite Plate Reader provides exceptional sensitivity and speed with minimal downtime - backed by our global network of application scientists and engineers. They can be used in a no-transfer protocol with suspension cells or with a transfer step for adherent cells. It features a modular design that allows users to add or change detection cartridges according to their needs. There are multiple factors to consider including: detection type, additional. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. PerkinElmer’s 96w low volume, 96w and 384w microplates can be read on all HTRF certified readers, thus making their implementation in your experiments really easy. The EnSight<sup>®</sup> multimode plate reader offers HTS Alpha and Ultra-sensitive Luminescence technologies alongside conventional multimode detection technologies. Push the screw down and turn it clockwise to lock the plate. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. For more. Taking these considerations into account, Cisbio’s 96w low volume and PKI’s 96w & 384w microplates have been carefully designed to provide the best signal detection environment possible to all HTRF assays. The Bradford assay was successfully performed on the BMG LABTECH microplate reader. Setu CLARIOstar readers can be equipped with monochromators or filters and only filter version is HTRF certified. Plate 25 μL of cells in. Enable more assay conditions Define individual reagents to be delivered to each well reducing reagent consumption and allowing for more assay conditions to be explored on a single microplate. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. The Z´ value takes into account four (4) parameters: the means (µ) and the standard deviations (σ) of the positive (plateau) and the negative (baseline) controls 1. 96-well culture plateFOR HTRF CERTIFIED READER. 96-well culture plate• HTRF®-Certified Reader. SpectraMax Paradigm Microplate Reader is a versatile and customizable device that can perform a wide range of detection modes, such as absorbance, fluorescence, luminescence, time-resolved fluorescence, and AlphaScreen. . pre+ 5 µL HTRFcompounds Activation Buffer. SpectraMax® M2e Multi-Mode Microplate Reader (top and bottom reading) MLDVM2E SpectraMax® M3 Multi-Mode Microplate Reader MLDVM3 Ordering information VWR cat. You can count on our plate readers to bring you the performance, reliability, consistent results, and the best user experience available—across a multitude of applications and a range of detection options. As shown in Fig. Read & analyse. Concentrate on running your assays instead of running your instrument: the optic modules contain all the filters, dichroics, and more needed to run your assay. Time-resolved fluorescence was measured on the SpectraMax Multi-Mode Microplate Readers (see Table 2 for instrument settings). The perfect microplate reader for everyday biochemical and cell-based assays, it measures at any wavelength, with top and bottom reading of all technologies. and TR-FRET, for example to detect PerkinElmer’s DELFIA, HTRF and LANCE assay technologies Time resolved fluorescence certified for use with HTRF. The reader only allows high performance for the used HTRF measurement when the HTRF is run with white plates. Place the imaging filter cube on top of the LED cube you installed, and use the hex wrench to screw it to the LED cube. It delivers even higher sensitivity for time-resolved fluorescence (TRF) applications, and the Enhanced Security software option includes. 0. For more ® information about HTRF compatible readers and for set-up recommendations, please. Its software interface is easy to learn and use, and lets you control the instrument from almost any device – perfect for multiuser labs. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. certified for use with HTRF. Cisbio’s 96-well low volume plates can be used with all HTRF assays, for example second messengers, phospho-proteins, cytokines and biomarkers. The SpectraMax i3x and iD5 readers can be equipped with an HTRF-certified detection cartridge or an HTRF-certified detection system, respectively. The kit includes everything you need: reagents, buffers and a microplate. Specialized filters allow for IR fluorescence detection ranging from 10-40 times more sensitive than conventional monochromator detection microplates can be read on all HTRF certified readers, thus making their implementation in your experiments really easy. Reagents were dispensed following Cisbio’s ligand binding protocol (Figure 3). Download the brochure to learn how this compact and robust device can handle various detection modes, plate formats, and software options. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. • HTRF cAMP HiRange Assay Kit (Cisbio Bioassays, France) • 96-well half area plates (white) (Greiner Bio-One, Germany) • 384-well small volume plates (white and black) (Greiner Bio-One, Germany) HTRF RCK The HTRF RCK is used to confirm the general suitability of a reader for HTRF assays, as well as set up and performance validation. On BMG LABTECH´s NEPHELOstar Plus, a laser diode (at 635 nm) with a highly collimated beam is used as high-intensity light source. FOR HTRF CERTIFIED READER For. The Negative control is used to check the non-specific signal. Remove the plate seal and read on an HTRF compatible reader. HTRF is a proprietary time-resolved fluoresce nce technology that overcomes• HTRF cAMP HiRange Assay Kit (Cisbio Bioassays, France) • 96-well half area plates (white) (Greiner Bio-One, Germany) • 384-well small volume plates (white and black) (Greiner Bio-One, Germany) HTRF RCK The HTRF RCK is used to confirm the general suitability of a reader for HTRF assays, as well as set up and performance validation. TWO-PLATE ASSAY PROTOCOL FOR SUSPENSION CELLS: 25 µL cells ONE-PLATE ASSAY PROTOCOL: Add *Depending on the cell lines used, the lysis volume should be optimized. High Speed Monochromator offers a typical wavelength accuracy of <0. Finding a plate reader that offers compatibility with microplates sold by multiple leading vendors can protect both the long-term cost and the performance of assays. Make sure the setup for Eu 3+ Cryptate is used. For more information. • Our fastest benchtop plate reader – uses two detectors to enable simultaneous, dual-wavelength reading • Well established for laser-based Alpha and TRF technologies, as well as ultrasensitive luminescence • Time-resolved fluorescence certified for use with HTRF ® technology • Software that can facilitate 21 CFR Part 11 compliance for• ®HTRF -Certified Reader(5) (1) IBMX is the most commonly used phosphodiesterase Pan-inhibitor. PerkinElmer. You can count on our plate readers to bring you the performance, reliability, consistent results, and the best user experience available—across a multitude of applications and a range of detection options. Certified SpectraMax® M5e-HTRF Readers The SpectraMax M5 e reader has the same perfor mance specifications as the M5 but is certified for use with Cisbio Bioassays’ HTRF (Homogeneous Time-Resolved Fluorescence) technology. The HTRF Reader Control Kit provides an easy and straightforward way to assess your reader’s overall performance and can facilitate the identification of potential limitations. FOR HTRF CERTIFIED READER. Transcreener® ADP2 FP assay. Homogeneous Time-Resolved Fluorescence (HTRF) • Certified for detection of Cisbio’s HTRF assays • GPCR assays (cAMP cell based, membrane based) • Kinases and ATPases • Cytokine Assays TUNE Multi-Mode • cAMP Quantitation • Cell Migration • Cell Viability and Proliferation • Cytotoxicity • DNA/RNA Quantitation (Fluorescent) Homogeneous Time-Resolved Fluorescence (HTRF) . It measures 96-well plates in less than 20 seconds and 1536-well formats in under one minute. For more information. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Do not pre-mix the d2 and Cryptate solutions prior to dispensing Make sure to use the set-up for Eu Cryptate. 8 µL cells + 4 µL compounds + 4 µL supplemented ® lysis buffer (4X) + 4 µL HTRF pre-mixed antibodies. Because of its homogeneous and fast nature, the assay is a preferred method to determine the protein concentration of samples. FOR ORE INFORMATION Europe and other ountries 33(0)466 96 0 U. HTRF TWO-PLATE ASSAY PROTOCOL FOR SUSPENSION CELLS: ONE-PLATE ASSAY PROTOCOL: overnight at RT * * Depending on cell lines used, volume of lysis should be optimized. + 4 µL HTRF pre-mixed antibodies. Both readers demonstrated their ability to perform the cAMP G s HiRange assay with results well within Cisbio’s required. 5. 14 seconds for a 384-well plate. FOR HTRF CERTIFIED READER. Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READER. Each well requires 10 μL of 4X lysis buffer /well. Plate 25 μL of cells in. Part Number# Product Description Number of Plates# 66PL96001 HTRF 96 well low volume plate 1 plate 66PL96005 HTRF 96 well low volume plate 5 platesThe assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. VICTOR Nivo Multimode Plate Reader. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Fluorescence polarization (FP) is a fluorescence-based detection method that is widely used to monitor molecular interactions in solution. FOR HTRF CERTIFIED READER culture plate to a 384-well small volume white plate. TO PREPARE TISSUE SAMPLES: Please refer to the Technical Note “Optimize Your HTRF® Cell Signaling Assays on Tissues” onHow to use Synergy microplate readers for HTRF detection, a versatile technology for drug discovery and life science research. February 23, 2010, WINOOSKI VT, USA - The Synergy TM H4 Hybrid Microplate Reader from BioTek Instruments is now HTRF® certified. No washing steps are required. Interaction with the industry and research communities is essential for innovating! For more than 30 years, Cisbio has been serving the needs of the clinical and drug discovery communities, delivering a world-class portfolio of assays designed to help find answers faster. Make sure the setup for Tb Cryptate is used. The kit includes everything you need: reagents, buffers and a microplate. For HTRF readings, a 620/40 nm filter (PerkinElmer 2100-5760) and a 665/7. Read & analyse. These plates can be read on all HTRF certified readers, thus making their implementation in. Evaluation of the performance of the Spark™ multimode reader for far red fluorescence emission. Cells in each image are identified by SoftMax Pro Software and cell-by-cellHTRF®-certified, DLReady, and SpectraMax® DuoLuc™-certified. For more information about HTRF. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. The EnVision XCite multimode plate reader provides exceptional sensitivity across all detection technologies, while keeping you flexible with monochromator option and full upgradeability. EnVision Nexus ultimode Plate Reader 2 Benefits • Provides the speed and sensitivity to boost your lab’s productivity with ultra-high throughput • Freely configurable and upgradable to adapt to your needs today and tomorrow • Fully automatable for walkaway convenience • Certified for use with HTRF® technology Key features BMG LABTECH plate readers certified for LanthaScreen detection include the PHERAstar FSX, CLARIOstar Plus and VANTAstar. • Our fastest benchtop plate reader – uses two detectors to enable simultaneous, dual-wavelength reading • Well established for laser-based Alpha and TRF technologies, as well as ultrasensitive luminescence • Time-resolved fluorescence certified for use with HTRF ® technology • Software that can facilitate 21 CFR Part 11 compliance for Choose from over 50 certified or compatible plate readers More than 50 certified or compatible instruments, and counting! Your lab might even already have one. Incubate 4h at RT. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. It features a modular design that allows users to add or change detection cartridges according to their needs. Incubate 2 h at RT. Its. Read & analyse. For more information about microplate recommendations, please visit our website • Store the kit at -16°C. The readers detect light signals that result from biological, chemical, biochemical, or physical reactions. For a list of HTRF-compatible readers and set-up recommendations, please visit our website • Small volume (SV) detection microplates - Use white plate only. The HTRF cellular assays were found to be at least 16-fold more sensitive than the Western Blot. These plates can be read on all HTRF certified readers, thus making their implementation in. Lysates are then transferred to the assay plate for the detection of total-BTK by HTRF. This DLReady-certified microplate reader can also come equipped with Simultaneous Dual Emission for BRET applications and can be further upgraded to a multi-mode reader. Make sure the plate reader is turnedon and then open the MikroWin software on the computer. Revvity. They can be used in a no-transfer protocol with suspension cells or with a transfer step for adherent cells. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. It utilises an ultra-fast UV/vis spectrometer or filters for absorbance as well as highly sensitive filters for all other detection modes. Plate 25 μL of cells in. At its core, Synergy H1 is a monochromator-based multi-mode microplate reader with a quadruple grating architecture for robust performance and flexibility. White 384-well low volume plate Make sure to use the set-up for Eu3+ Cryptate. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. 27 / Pack of 100; Qty. For more information about HTRF. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. The kit includes everything you need: reagents, buffers and a microplate. Plate 25 μL of cells in. For more information about set-up and compatible HTRF ® readers,• HTRF®-Certified Reader. Other technologies such as imaging. 1 – 1. PLATE READER INTRODUCTION Selecting the right plate reader for your lab can be challenging. n HTRF plate validated (QC with reference HTRF assays) n Low to high volumes (10 µL to 350 µL) n Can be used with all HTRF compatible readers n For cell-based or biochemical assays Trying is Believing! Plate Characteristics HTRF® 96 - well low volume plate CulturPlateTM 96 - well Microplate CulturPlate TM 384 - well Microplate ProxiPlateTM 384 - Cisbio’s 96-well low volume plates can be used with all HTRF assays, for example second messengers, phospho-proteins, cytokines and biomarkers. Figure 1: Principle of HTRF Cortisol competitive assay. Incubate 2h at RT Transfer 16 µL supernatant. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. If not, our scientific team will help you choose the instrument that best suits your needs and budget. HTRF-specific. FOR HTRF CERTIFIED READER. HTRF® CertifiedCertified for use with HTRF ®, Alpha, LANCE ® or DELFIA ®. After an overnight incubation, the plate is washed 3 times. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. 1) . Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. If not, our scientific team will help you choose the instrument that best suits your needs and budget. When equipped with a quad monochromator, the EnSight plate reader can improve performance for absorbance and fluorescence detection. Read & analyse. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. HTRF-specific. The CLARIOstar® Plus is our most flexible multi-mode plate reader. HTRF is a proprietary time-resolved fluoresce nce technology that overcomes To prevent evaporation during assay incubation steps, we recommend the use of TopSeal-A PLUS, a clear adhesive seal for microplate. Plus . The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. Preconfigured HTRF. Certified for use with HTRF ®, Alpha, LANCE ® or DELFIA ®. Incubate overnight at RT. Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READERFOR HTRF CERTIFIED READER. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. Read & analyse. Cisbio’s 96-well low volume plates can be used with all HTRF assays, for example second messengers, phospho-proteins, cytokines and biomarkers. Transcreener® ADP2 TR-FRET assay. 1, introducing a time delay of approximately 50 to 150 µ seconds between the system excitation and fluorescence measurement allows the signal to be cleared of all non-specific short-lived emissions. ONE-PLATE ASSAY PROTOCOL: Small volume detection plate. DLReady™ certified luminometer plate readers. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. At that stage, the analyte is sandwiched between the 2 antibodies. Not for use in diagnostic procedures. CLARIOstar is equipped with a specific optical device which enables the measurement of. The multidetection microplate readers CLARIOstar, PHERAstar FS, FLUOstar Omega, and POLARstar Omega are all HTRF-certified by Cisbio, meaning each of the. For more. CLARIOstar is equipped with a specific optical device which enables the measurement of both 620 nm cryptate and 665 nm acceptor emissions. Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. Combined with our Kaleido™ workflow software, it. FOR HTRF CERTIFIED READER. Lysates are then transferred to the assay plate for the detection of Phospho-AKT-3 (Ser473) by HTRF. For instance, when assay volumes are. The assay can be run under a two-plate assay protocol, where cells are plated, stimulated and lysed in the same culture plate. spectramax paradigm system, htrf reagents, spectramax i3 and paradigm readers, HTRF assay, htrf certifiedLysates are then transferred ®to the assay plate for the detection of Total CHK2 by HTRF reagents. Partnerships for delivering turn-key solutions. For more information. This protocol gives the cells viability and confluence to be monitored. The plate map included a low and a high level calibrator, conjugate blanks, and a specific control to assess the counting rate at 620 nm (i. • Clone your plate data for multiple data reduction optionsExpanded read mode capabilities Equipped for high performance, the SpectraMax iD5 is HTRF®-certified, DLReady, and SpectraMax® DuoLuc™- certified. For more ® information about HTRF compatible readers and for set-up recommendations, please. Its. HTRF is a proprietary time-resolved fluorescence technology that overcomes Results summary for HTRF IP1 standard curve. Read & analyse. Incubate overnight at RT. For more. Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READER. + 4 µL HTRF ® pre-mixed antibo dies 2. The PHERAstar ® FSX and CLARIOstar ® microplate readers offer the best possible performance for AlphaScreen ®, AlphaLISA ®, and AlphaPlex™ assays. The PHERAstar ® FSX is our most sensitive multi-mode reader for high-throughput screening, combining this sensitivity with fast read times. Lysates are then transferred to the assay plate for the detection of Total Cofilin by HTRF® reagents. These are the different kinetic modes on BMG LABTECH microplate readers. Plus . Example assays are ELISAs, DNA and protein concentration determination, steady-state dose response curves, ADP-Glo™ (Promega), Transcreener™ GDP fluorescent polarization. For more information about HTRF. Cisbio’s 96-well low volume plates can be used with all HTRF assays, for example second messengers, phospho-proteins, cytokines and biomarkers. Suggested plate map: 1: 2 3: A: 14 μL Std 0: 3 μL IP1 d2 Reagent 3 μL IP1 Tb Cryptate Antibody• HTRF®-Certified Reader. For more information about HTRF ® compatible readers and for set-up recommendations, please. Lysates are then transferred to the assay plate for the detection of total-RET by HTRF. 1, introducing a time delay of approximately 50 to 150 µ seconds between the system excitation and fluorescence measurement allows the signal to be cleared of all non. The mean and standard deviation. After defining the objectives and their locations in Gen5: 1. This protocol gives the cells viability and confluence to be monitored. Expanded read mode capabilities Equipped for high performance, the SpectraMax iD5 is HTRF®-certified, DLReady, and SpectraMax® DuoLuc™- certified. Plate Type and Read Area should be selected based on the microplate used and wells to be read. The new ligands are divided into four groups according to their chemical structures and/or interactions with YTHDC1 (Table 1 and Fig. Step 5 Remove the plate sealer and read on an HTRF® compatible reader The blank control is used to check the Cryptate signal at 620 nm. For instance, when assay volumes are. HTRF combines time-resolved fresh detection with FRET and is mainly used for drug screening in high-throughput. 96-well or 384-well small volume (SV) detection microplates - For more information about microplate recommendations, please. This protocol, HTS designed, allows. Depending on cell lines used, it can be necessary to dilute the cell lysate to ensure samples are within the assay linear range FOR HTRF CERTIFIED READER. Lysates are then transferred to the assay plate for the detection of total-p53 by HTRF. FOR HTRF CERTIFIED READER. Plate 25 μL of cells in. Discover our high performance reader. Read & analyse. Specialized microplate readers are available that enable HTRF (Homogeneous Time-Resolved Fluorescence) experiments and analyses. FOR HTRF CERTIFIED READER. use of a wide variety of plates or sample matrix formats. Sensitive, accurate and reliable detection is critical across cell, protein and genomic analysis applications in drug discovery and development. FOR HTRF CERTIFIED READER. com HTRF Microplate Readers. FOR HTRF CERTIFIED READER For more ® information about HTRF compatible readers and for set-up recommendations, please visit our. FOR HTRF CERTIFIED READER. The VICTOR Nivo. • HTRF plate validated (QC with reference HTRF assays) • Low to high volumes (10 µL to 350 µL) • Can be used with all HTRF compatible readersONE-PLATE ASSAY PROTOCOL: Small volume detection plate. Lysates are then transferred to the assay plate for the detection of phospho-Tie2 (Tyr992) by HTRF. Certified for use with Alpha, LANCE ® or DELFIA ®. Synergy NEO2 readers must be appropriately configured for HTRF® readout by setting up the measurement conditions in the Gen5™ Reader Control and Data Analysis Software. At Cisbio we provide microplates that help you discover better. PHERAstar FSX Powerful and most sensitive HTS plate reader . The dual measurement. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. PerkinElmer offers two microplate readers certified for use with HTRF: Envision. It also integrates with the. 55 µL anti-cAMP-Cryptate. $352. Certified for use with HTRF ®, Alpha, LANCE ® or DELFIA ®. 47 seconds for a 1536-well plate. HTRF combines standard FRET technology with time-resolved measurement of fluorescence, eliminating short-lived background fluorescence. $1,024. According to the manufacturers protocol, this protein assay is linear in the range of 0. For the standard curve described, all wells of the microplate are read. The Spark multimode reader from Tecan performs accurate and sensitive measurements of fluorescent cell-based assays. • ®HTRF -Certified Reader(5) (1) IBMX is the most commonly used phosphodiesterase Pan-inhibitor. They can be used in a no-transfer protocol with suspension cells or with a transfer step for adherent cells. Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. Customer Transformation Hub. culture plate. Seal the plate and incubate 1 hour at RT : Remove the plate sealer and read on an HTRF® compatible reader . Add Lysis Add * Depending on cell lines used, volume of lysis should be optimized. The plate is set up for reading and images are acquired according to specified parameters. Due to the chemical and photophysical. HTRF TWO-PLATE ASSAY PROTOCOL FOR SUSPENSION CELLS: ONE-PLATE ASSAY PROTOCOL: overnight * * Depending on cell lines used, volume of lysis should be optimized. Lysates are then transferred to the assay plate for the detection of total-Tau by HTRF. FLUOstar Omega Filter-based multi-mode microplate reader . Page 30 4. PROTOCOL AT A GLANCE. FOR HTRF CERTIFIED READER. The DLR assay is a two step reaction that uses two luciferase enzymes, Firefly and Renilla (Figure 1). Plate 50 µL of cells in 96-well tissue-culture treated plate in appropriate growth medium and incubate overnight, at 37°C in CO2 atmosphere. Multimode plate readers include, at the very least, some combination of absorbance (typically covering the UV/visible range), fluorescence and luminescence. HTRF® Europium cryptate donor / Red acceptor readout Setup recommendations for Tristar 5FOR HTRF CERTIFIED READER. View Brochure. 5 EXAMPLE OF MAP PLATE . (In the following text, the term YTHDC1 is used frequently as an abbreviation for the m 6 A-recognition domain of YTHDC1). Make sure the setup for Tb Cryptate is used. FOR HTRF CERTIFIED. Excitation lasers significantly improve performance and lower. Your lab might even have one already. • HTRF plate validated (QC with reference HTRF assays) • Low to high volumes (10 µL to 350 µL) • Can be used with all HTRF compatible readersFOR HTRF CERTIFIED READER. The plate is set up for reading and images are acquired according to specified parameters. Plate 25 μL of cells in 96 half-well plate in your appropriate medium. Figure 1: Principle of HTRF Prostaglandin E2 competitive assay. Cisbio’s 96-well low volume plates can be used with all HTRF assays, for example second messengers, phospho-proteins, cytokines and biomarkers. Each well generally requires 50µL of lysis buffer 1X. You can verify your reader’s ability to detect and use HTRF technology, then easily configure your instrument with its own HTRF setup. Plate 25 μL of cells in.