ArrayIt® has developed a sophisticated microarray substrate coated with a hydrophobic polymer for a wide spectrum of protein microarray applications. The 150 µm thick layer of hydrophobic polymer is bound to an atomically flat glass substrate slide for high precision and uniformity. The ultimate surface for protein microarray applications provides small printed spots, strong signals, and ultra-low background. Use in conjunction with the SpotWare Colorimtric Scanner and SuperNitro Microarray Substrates to transform your traditional filter assays into a microarray format for highly affordable research and in vitro diagnostics.
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Product Description The SuperProtein surface provides a white micro porous polymer laminated to a SuperClean substrate at an affordable price. The surface binds proteins via efficient hydrophobic interactions. This unique hydrophobic surface is perfect for spotting multiple samples, multiple times over multiple substrates with 1 low volume loading of sample using the 946 Micro Spotting Device. SuperProtein Substrates are manufactured in class 1 cleanrooms. Cleanroom manufacture eliminates contamination of the microarray surface with particulates, proteases, nucleases and other contaminants that impair the quality of microarray experimentation.
SuperProtein can be used for the following protein microarray applications:
Protein – Protein
Protein – Antibody
Protein – Antigen
Protein – DNA
Protein – RNA
Protein – Drug
Protein – Carbohydrate
ELISA in a microarray format
Multiplexing Reaction Tools SuperPVDF slides are compatible with the multiwell reaction tools, AHC1x16, AHC1x24, AHC4x24, and MMH4x24. Using these tools and the right microarrayer programming it is possible to run up to 24 separate microarray experiments on a single slide.
Users will appreciate the following features of SuperProtein Substrates:
Manufactured in state-of-the-art class 1 microarray cleanrooms
Free of particulate, protease and nuclease contamination
Standard 1” x 3” size (76 x 25 x 1.2 mm including hydrophobic polymer)
Corner chamfer (upper right corner) for unambiguous orientation
High efficiency hydrophobic protein coupling
Supports both contact printing and ink-jet printing
No cross-linking or baking required for coupling
“Zero” background fluorescence when used with the SpotWare Scanner
Binds proteins, enzymes, antibodies, receptors, antigens, and more…
Print pure proteins, recombinant proteins and cellular extracts
Uniform feature size
25 substrates per box
Anti-static and impact resistant packaging
1 year shelf life at room temperature
Assay for Detection of Antibodies to Target Antigens
Figure 1. Antigens are printed on SuperProtein surface. Patient serum containing measured antibody is incubated to the microarray. After reaction a secondary antibody containing enzyme linked conjugate is incubated and subsequently developed to allow colorimetric detection of the measured antibody. The use of an incorporated internal calibration curve for each subclass of antigen/antibody on the microarray provides a unique advantage over current immunoassays protocols, where the standard curve and the sample are processed in separate tubes or wells.
Figure 2. Correct Substrate Orientation. Shown is a graphic of two ArrayIt® Microarray Substrates, 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® Microarray Substrates is suitable for printing. Please print on the correct side only.
Complete Protocol: 1. Print purified proteins suspended in PBS at 0.1-1 µg/µl as a final concentration from an MMP384 microplate with no more than 15 µl per well with a SpotBot Protein Edition Microarrayer, NanoPrint Protein Microarrayer or compatible device. A 50kD protein at 1 µg/µl concentration has a concentration of 20 µM. Assuming a 30% coupling efficiency, a 20 µM protein will produce a target density of 1011 proteins per mm2 of substrate. Proteins retain binding and enzymatic activity on the surface. Printed microarrays should be stored unprocessed at 4C to protect coupled molecules. Processing should be performed just prior to use for best performance.
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 room temperature is usually sufficient to obtain a strong signal. Reactions are also performed by pipetting 10 L defined volume of sample or control (primary antibody) into Petri dish in 10 ml of blocking buffer.
4. Appropriately discard the reaction sample or control solutions and wash the slides in 1X PBS, 3 times at 10 minutes each in a High Throughput Wash Station.
5. React microarray with an enzyme conjugated labeled 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 60-minute incubation at room temperature is usually sufficient to obtain a strong signal.
7. Use appropriate enzymatic developer. For quantitative work, pre-determine development time by conducting pilot study. It is important that all microarrays to be compared are developed by the same time procedure. It is not always necessary to see signal by eye, the SpotWare Colorimetric Microarray Scanner can detect signals that are invisible to the naked eye.