Arrayit SuperProtein Microarray Substrate Slides Barcoded (Box of 25) provide atomically smooth glass substrate slides coated with a 150 µm layer of SuperProtein membrane. SuperProtein substrate slides support the standard glass slide format (1 x 25 x 76 mm) and have a corner chamfer to allow unambiguous orientation during use. A time-tested membrane surface for a wide variety of fluorescent and colorimetric DNA and protein microarray applications, provides small printed spots, strong and uniform signals, and low background. Use in conjunction with the colorimetric SpotWare scanner and Arrayit's extensive line of microarray products and services, 25 slides per box with barcodes.
Microarray Substrates and Slides - SuperProtein Membrane Coated Microarray Substrate Slides for DNA and Protein Microarray Manufacturing
Arrayit has developed an advanced microarray substrate slide product line featuring a proprietary hydrophobic polymer coating for a wide spectrum of protein and DNA microarray applications. The 150 µm thick layer of hydrophobic polymer is bound to atomically smooth glass substrate slides for high precision and uniformity. SuperProtein is the ultimate surface for protein and DNA microarray applications, providing small printed spots, strong signals, and ultra-low background. Use in conjunction with the SpotWare Colorimetric Scanner and SuperNitro Microarray Substrate Slides to transform your traditional filter assays into a microarray format for highly affordable research, life sciences and microarray testing applications.
Table of Contents
- Product Description
- Technical Information
- Short Protocol
- Complete Protocol
- Technical Support
- Scientific Publications
- Ordering Information
- Warranty
Product Description
Arrayit SuperProtein Microarray Substrate Slides feature a white micro porous polymer surface laminated to SuperClean substrate slides to empower affordable colorimetric microarrays. The surface binds proteins and DNA via efficient hydrophobic interactions. This unique hydrophobic surface is ideal for printing multiple samples, multiple times over multiple substrate slides with one low volume sample using Arrayit microarray printing technology. Arrayit SuperProtein Substrate Slides are manufactured in a cleanroom environment, which eliminates contamination of the microarray surface with particulates, proteases, nucleases and other contaminants that impair the quality of microarray experimentation.
Users will appreciate the following features of Arrayit SuperProtein:
- Manufactured in microarray cleanrooms
- Free of particulate, protease and nuclease contamination
- Standard 1.2 x 25 x 76 mm dimensions including hydrophobic polymer
- Corner chamfer in the upper right corner enables unambiguous orientation
- High efficiency hydrophobic protein and DNA coupling
- Supports both contact printing and ink-jet printing
- No cross-linking or baking required for coupling
- Low background fluorescence when used with Arrayit SpotWare Colorimetric Microarray Scanners
- Binds proteins, enzymes, antibodies, receptors, antigens and DNA
- Print pure proteins, recombinant proteins and cellular extracts
- Uniform printed feature size
- 25 substrate slides per box
- Anti-static and impact resistant packaging
- 1 year shelf life at room temperature
Technical Information
Arrayit SuperProtein can be used for the following protein microarray molecular interactions and applications:
- protein-protein
- protein-antibody
- protein-antigen
- protein-DNA
- protein-RNA
- protein-drug
- protein-carbohydrate
- converting enzyme-linked immunosorptive assays (ELISA) into a microarray format
Multiplexing Reaction Tools:
Arrayit SuperProtein slides are compatible with the multiwell reaction tools AHC1X16, AHC1X24, AHC4X24, and MMH4X24. Arrayit hybridization cassettes, coupled with the proper microarrayer programming, enable customers to process up to 24 separate microarray specimens per slide and up to 96 specimens per hybridization cassette for high-throughput research and microarray testing applications.
Example Protocol:
See Figure 1 as an example assay schematic for the detection of reactive antibodies using an antigen microarray
Figure 1. Antigens are printed on an Arrayit SuperProtein microarray substrate slide. Patient serum containing presumptive reactive antibodies is incubated to the microarray. After the primary binding reaction, a secondary antibody containing an enzyme-linked conjugate or fluorescent tag is incubated and subsequently developed to allow colorimetric detection of the antibodies in the specimen. An internal calibration curve for each subclass of antibodies detected on the microarray provides a unique advantage over traditional immunoassays, where the standard curve and the specimen are processed in separate tubes or wells.
Figure 2. Correct substrate slide orientation. Shown is a graphic of two Arrayit SuperProtein Microarray Substrate Slides, showing the correct and incorrect orientation for use. In the correct orientation (blue graphic), the chamfer will be located in the upper right corner and samples should be printed on the side facing upward, which is the same side that contains the word “Correct!”. In the incorrect orientation (red graphic), the chamfer will be located in the upper left corner, placing the backside facing upward, which is the side that contains the word “Incorrect!”. Only one side of Arrayit SuperProtein Microarray Substrate Slides is suitable for printing. Please print on the membrane side only.
Figure 3. Arrayit SuperProtein Microarray Substrate Slides can be purchased with unique barcode identifiers to facilitate sample tracking during manufacturing and use.
Short Protocol:
- Print antigens into a microarray using a an Arrayit protein edition microarray printer.
- Wash and block using Arrayit protein microarray buffers.
- Perform binding reaction with patient research specimen.
- Wash with Arrayit protein microarray wash buffer (Cat. PMWB).
- Stain the microarray with a secondary labeled antibody.
- Wash.
- Develop by enzymatic reaction (or use direct fluorescence).
- Scan with Arrayit SpotWare Colorimetric Scanner and analyze the microarray data.
Complete Protocol:
1. Print purified proteins suspended in Arrayit Protein Printing Buffer at 0.25-0.5 µg/µl using an Arrayit MMP384 384-well microplate with no more than 12.5 µl per well with an Arrayit protein microarray printer. A 50kD protein at 0.5 µg/µl concentration has a concentration of 10 µM. Assuming a 30% coupling efficiency, a 10 µM protein will produce a target density of 500 proteins per mm2 of substrate slide. Proteins retain binding and enzymatic activity on the surface. Printed microarrays should be stored unprocessed at 4°C to protect coupled molecules. Processing should be performed just prior to use for best performance.
2. Wash the printed microarrays by adding 500 ml of Protein Microarray Wash Buffer (Cat. PMWB) in a High Throughput Wash Station and wash with gentle agitation for 5 minutes. Remove printed microarray and spin dry for 2 seconds using a Microarray Centrifuge. Block using 4 µl for every square centimeter of coverslip with SuperProtein blocking buffer for 15 minutes and then wash for 5 minutes with Protein Microarray Wash Buffer using a High Throughput Wash Station and spin dry.
3. Perform binding reactions in blocking buffer plus primary antibody or control sample diluted 1:1,000. Samples can be incubated as a droplet on the printed microarray, underneath a cover slip, or in a micro-fluidics chamber. A 60-minute incubation at 25-37°C is usually sufficient to obtain a strong signal. Reactions are also performed by pipetting 10 µl of sample or control (primary antibody) into a Petri dish in 10 ml of blocking buffer.
4. Discard the reaction sample or control solutions and wash the slides in Protein Microarray Wash Buffer 3 times for 5 minutes per wash in a High Throughput Wash Station.
5. React microarray with an enzyme-conjugated secondary antibody for 1 hour (e.g. goat anti-human IgG-AP). Use a dilution of 1:1000. Samples can be incubated as a droplet on the printed microarray, underneath a cover slip, or in a micro-fluidics chamber. A 30-minute incubation at 25-37°C is usually sufficient to obtain a strong signal.
6. Wash away secondary antibody using High Throughput Wash Station and Protein Microarray Wash Buffer (Cat. PMWB) 3 times for 5 minutes each.
7. Use appropriate enzymatic developer. For quantitative work, pre-determine development time by conducting a pilot study. It is important that all microarrays are developed using the same ampount of time. It is not always necessary to see signal by eye, as the SpotWare Colorimetric Microarray Scanner can detect signals that are invisible to the naked eye.
8. To halt assay color development, immerse the microarrays in distilled water 1 minute. Spin the slides dry in an Arrayit Microarray Centrifuge. Microarrays are now ready to scan in the SpotWare Colorimetric Microarray Scanner.
Recommended Equipment and Reagents
NanoPrint™ 2 Microarrayers
NanoPrint™ 2 Protein Edition Microarrayers
SpotBot® 4 Personal Microarrayers
SpotBot® 4 Protein Edition Personal Microarrayers
SuperNitro Microarray Substrate Slides
SuperProtein Microarray Substrate Slides
Protein Printing Buffer
BlockIt™ Microarray Blocking Buffer
SuperProtein Blocking Buffer
PCR Purification Kits
Arrayit Green and Red Reactive Fluorescent Dyes
Microarray High-Speed Centrifuge
High-Throughput Wash Stations