Q-Plex: Cytokines multiplexed ELISA based test - Per risparmiare Tempo, Campioni e Denaro

Recombinant Cytokines and Growth Factors Cytokines and Growth Factors ELISA Kits and Development kits   Tecniche a Confronto:      Electro-Chemiluminescent Bead Based Q-Plex multiplex ELISA Time 864 data points / test 1000 data points / test 1536 data points / test Sample Size 50-100 l / test 12.5-50 l / test 5-30 l / test Cost $1,920 (10-Plex) $1,915 (9-Plex) $1,090 (9-Plex) Protocol Complex-electronic chip  in 96 well plate Complex-microfluidic channels Similar to ELISA Equipment Equipment ($100-200K) Equipment ($100-200K) Q-View Imager ($10K) Sample Types Many types Difficulty with Serum and other viscous samples Many types Variation Acceptable Variability ≤10% CV Bead-Lot to Lot Variability Low Variability ≤7% CV Cost Per Data Point $2.00 per data point $2.02 per data point $1.00 per data point Sensitivity Pg/ml Pg/ml Pg/ml       Human Euro         150251HU Q-Plex Human Angiogenesis (9-plex) 1.155 ANG-2, FGF basic, HGF, IL-8, PDGF-BB, TIMP-1, TIMP-2, TNFα, VEGF        120251HU Q-Plex Human Chemokine (9-plex) 1.155 Eotaxin, GROa, I-309, IL-8, IP-10, MCP-1, MCP-2, RANTES, TARC        110451HU Q-Plex Human Cytokine - Inflammation (9-plex) 1.155 IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IFNγ, TNFα        130251HU Q-Plex Human Cytokine - IR (9-plex) 1.155 IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12p70, IFNγ, TNFα        110351HU Q-Plex Human Cytokine - Stripwells (16-plex) 1.691 IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-23, IFNγ, TNFα, TNFβ        110951HU Q-Plex Human Cytokine - Screen (16-plex) 1.578 IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-23, IFNγ, TNFα, TNFβ        130951HU Q-Plex Human Cytokine - Screen IR (16-plex) 1.578 IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IFNγ, MCP-1, TNFα, TNFβ        Mouse            130251MS Q-Plex Mouse Cytokine - IR (9-plex) 1.155 IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12p70, IFNγ, TNFα        130951MS Q-Plex Mouse Cytokine - Screen IR (16-plex) 1.578 IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-17, MCP-1, IFNγ, TNFα, MIP-1α, GMCSF, RANTES        110351MS Q-Plex Mouse Cytokine - Stripwells (16-plex) 1.691 IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-17, MCP-1, IFNγ, TNFα, MIP-1α, GMCSF, RANTES        110951MS Q-Plex Mouse Cytokine - Screen (16-plex) 1.578 IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-17, MCP-1, IFNγ, TNFα, MIP-1α, GMCSF, RANTES        110451MS Q-Plex Mouse Cytokine - Inflammation (14-plex) 1.480 IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-6, IL-10, IL-12p70, IL-17, MCP-1, TNFα, MIP-1α, GMCSF, RANTES        Manuali: Human Cytokine - Stripwells (16 plex) Human Cytokine - Inflammation (9-plex) Human Cytokine - Screen (16-plex) Human Angiogensis (9-plex) Human Cytokine - IR (9-plex) Human Cytokine - Screen IR (16-plex)   Mouse Cytokine - Screen (16-plex) Mouse Cytokine - Inflammation (14-plex) Mouse Cytokine - IR (9-plex) Mouse Cytokine - Screen IR (16-plex)   Q-View Imager Q-View Software

SUPPORT FILES FAQ Imagers Q-View Imager Alpha Innotech HD2 Bio-Rad Versa Doc 4000 Bio-Rad XRS LI-COR Aerius LI-COR Odyssey SearchLight Imager Fujifilm LAS - 3000 Kodak 4000 mm Alpha Innotech Fluorchem SP UVP EC3 Darkroom Literature Posters Product Information Sheets Technical Briefs Manuals Videos Alpha Innotech FC2 for Multiplex Elisa Alpha Innotech HD2 for Multiplex Elisa Bio Rad Chemi Doc XRS for multiplex elisa Bio Rad Versa Doc for multiplex elisa Fuji Film LAS 3000 for Multiplex Elisas Licor Odessey Setup for Multiplex Elisas Quansys Chemi Assay for Multiplex Elisas Quansys Q-View Imager Setup for multiplex elisas Quality Applications FAQ How does this array work? The Q-Plex technology consists of a printed array of capture antibodies to a variety of biological markers. The user adds an antigen standard and samples to individual wells of a 96-well plate. After incubation and washing, a mixture of biotinylated antibodies is added to the plate. This is followed by incubating with streptavidin-HRP or -dye and is completed by capturing the light produced from adding chemiluminescent substrate with an imaging system or fluorescent scanner such as the LI-COR Odyssey. Pixel intensity values from the images are used to construct a standard curve and calculate the concentrations of the unknown sample for each system. Please visit this webpage for a detailed description of our technology.   Are the results quantitative? Yes, the array uses a standard curve to accurately quantify samples and the curve has been designed to encompass the biological ranges for individual proteins. Does the standard curve encompass the lower limit of detection (LLD) for each cytokine? The LLD is the lowest concentration of an analyte where its signal is distinguishable from the background (2 times the standard deviation of the mean of the background). This is different than the lower limit of quantification (LLOQ) which is the lowest concentration of an analyte that can be accurately measured. In many cases the typical standard curve does not reach the LLD nor the LLOQ for a given protein. The user has an option of using additional dilutions of the antigen mix to achieve the lower limit of quantification (LLOQ) for a given protein. A quick determination of LLOQ is to examine the backfit of your regression analysis and determine where the model no longer predicts within 80-120% of the actual standard value. I.E. the LLOQ is the lowest point where the backfit is within 80-120% of its corresponding value on the standard curve. How should I dilute my sample if I expect a high response for one bio marker and a low response for another? We recommend testing each sample in step dilutions. For example, when testing cell culture supernatant for unknown cytokine concentrations; a 1:2, a 1:10, and a 1:50 dilution are typically used. Will different CCD and CMOS cameras give different pixel intensities and how will this affect my assay? Yes, different CCD cameras give different pixel intensities based on the sensitivity of the camera. However, this will not affect assay performance or standard curve generation as the pixel intensities are relative values. Do I have to use the entire 96 well plate in one assay? Once the lyophilized reagents are reconstituted they should be used immediately. If the user desires to run several assays using the 96 well plate, we recommend the user purchase the Q-Plex Stripwell kits. These kits come with additional reagents for multiple testing periods. However, customers can use regular retail kits as long as the contents are stored properly. The reconstituted antigen standards should be immediately aliquoted and frozen at -20C. The entire kit contents should be used within one week of reconstitution. Detailed information on kit storage after reconstitution can be found in the Users manual. Can I stop the assay once I start it? No, for best results it is recommended that the assay be completed once started. After reading the results with a camera, the user has the option to delay data analysis to a convenient time. The IR plate may be read at any time after completion of the assay. What additional equipment (not provided in the kit) are required to run this assay? The user will need to provide a pipette and imager. We also recommend an automatic plate washer and multichannel pipette for best results. What imagers can be used to image the Quansys Multiplex ELISA plate? TThe Q-Plex Array plate can be read by a number of imaging systems and gel documentation systems such as Alpha Innotech Fluorchem SP, HD2, and FC2, Kodak 4000 MM, Fujifilm LAS-3000 and LAS-4000, Bio-Rad Versa Doc 4000 and ChemiDoc XRS, and others. Please visit this webpage for a complete listing of our recommended imagers. A cooled and non-cooled CCD imager with at least 12-bit outputs contained in a light tight box are appropriate for imaging. Our array format and the Q-View Software is compatible with most imaging systems. High-resolution cameras in light tight boxes with at least 19″ from the lens to the plate if imaging from the top or 6″ from the bottom are ideal for this application, but not required. Please call technical support for any imager related questions (1-888-782-6797 ext 321). How is the data gathered from the CCD imager analyzed? The Q-View Software allows the user to open the plate image, mark the spots, input well assignements and select the product type. The software then automatically analyzes the pixel intensity of individual spots on the plate and process the data. All customers can download a free trial of the software and have full access for 20 days. After 20 days, the software becomes the trial version which will still process images and calculate raw data but will not perform regression analysis. Please contact us for more information on Q-View Software or visit this webpage. What kind of samples can be run in the Quansys Multiplex ELISA Array? The Q-Plex Array has been tested with serum, plasma, urine, and cell and tissue lysate. We have experienced no interference from FBS, 1% NP-40, 1% Tween, 1% Triton X-100, heparin (30 mg/ml), 1M urea, 20 mM EDTA, 20% citrate. SDS has been shown to interfere with the assay. Human samples containing human antibodies must be diluted 1:2 in the provided sample dilution buffer to remove potential false postives. If tissue homogenates or cell lysates are used, centrifuging is recommended to clarify samples prior to using in the assay. When using tissue homogenates, we recommend preparation in a protein-free buffer so that samples can be tested for total protein to normalize cytokine responses. How much sample is required for the Quansys Multiplex ELISA Array? The Q-Plex Array requires 30 l total volume per well (sample plus diluent). Smaller sample volumes may be used when high levels of detectable proteins are anticipated. Samples containing antibodies MUST be diluted 1:2 in sample dilution buffer prior to running the assay. How long does it take to complete the Quansys Multiplex ELISA protocol? Most protocols require as little as two and a half hours for completion including analysis with the Q-View Software. How much variance does the Quansys Multiplex ELISA array have? The Quansys Multiplex ELISA assay has <15% CV between kit lots. Samples are usually linear out to a 1 to 64 dilution. I want to measure a protein I dont see displayed or I need to measure a unique set of cytokines that you dont currently offer. Can Quansys build an assay for me? Yes, Quansys Biosciences specializes in assay development, please call or email for details and pricing. I dont have access to a CCD camera system. Is there a testing service? Yes, your samples can be mailed to Quansys where they will be tested using the Quansys Multiplex ELISA platform. Data, along with statistical analyses will be provided to the user within two weeks. Please call for shipping details and pricing.

Imaging Systems Recommended for Quansys Biosciences Antibody Arrays

The following is a list of imaging systems compatible for use with Quansys Biosciences Q-Plex multiplex ELISA technology.

Top Performance

• * Q-View Imager and Software.

Engineered for minimal optical distortion and automated via Q-View Software.  This is recommended for the acquisition of the chemiluminescent signal and quantification of spot intensity for the Q-Plex multiplex ELISA arrays.

Recommended

• * Alpha Innotech: HD2 and FC2
• * Bio-Rad: Versa Doc 4000 and XRS
• * LI-COR: Aerius, Odyssey
• * SearchLight Imager
• * Fujifilm: LAS-3000, LAS-3000 Mini, and LAS-4000.
• * Kodak: 4000MM

These Imagers have built in correction for image distortions such as vignetting and parallax via mechanical or software applications.  Theyve all been tested extensively with the Q-Plex products and have provided accurate and reproducible data.

Suitable

• * Alpha Innotech: Fluorchem HD, SP, 8000, 8900, 9900

These suitable imagers can have inherent variation from 9-12% within the field of view.  Alternative imagers can be used for quantitative analysis dependant upon the accuracy required.

Qualitative

• * UVP: BioDoc-IT System, EC3 Darkroom
• * Kodak: 2000MM, Gel Logic 100

Due to low resolution or non cooled cameras, quantitative abilities of these imagers can be questionable.  However, for qualitative applications, the alternative imagers are suitable.