Arrayit SuperPVDF Microarray Substrate Slides (Box of 5) provide atomically smooth glass substrate slides coated with a 150 µm layer of polyvinylidene fluoride (PVDF). SuperPVDF 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, 5 slides per box.
Microarray Substrates & Slides - SuperPVDF Microarray Substrate Slides with Polyvinylidene Fluoride (PVDF) Coating for DNA and Protein Microarray Manufacturing
Arrayit provides an advanced microarray substrate slide product line featuring a polyvinylidene fluoride (PVDF) membrane coating for DNA and protein microarray applications. The 150 µm thick layer of PVDF is bound to atomically smooth glass substrate slides (1 x 25 x 76 mm) for high precision and uniformity. PVDF is an excellent surface for a variety of microarray applications, providing small printed spots on a highly hydrophobic surface. When used in conjunction with the SpotWare Colorimetric Scanner, background is extremely low. SuperPVDF makes it possible to transform traditional PVDF filter dot blot assays into a microarray format for highly affordable research, life sciences and microarray testing applications.
Note: Arrayit PVDF Microarray Substrate Slides are compatible with Arrayit hybridization cassettes to multiplex the number of microarrays per PVDF substrate slide.
Table of Contents
- Product Description
- Technical Information
- Short Protocol
- Complete Protocol
- Technical Support
- Scientific Publications
- Ordering Information
- Warranty
Product Description
Arrayit SuperPVDF Microarray Substrate Slides provide a white micro porous polymer surface laminated to SuperClean substrate slides at an affordable price. This surface binds proteins and other biomolecules through efficient hydrophobic interactions based on Van der Waals forces. Microarrays can manufactured using microarray printers running Arrayit microarray printing technology, and other types of printers can be used as well. SuperPVDF Substrate Slides are manufactured in a cleanroom environment to eliminate 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 SuperPVDF Substrate Slides:
- Manufactured in microarray cleanrooms
- Free of particulate, protease and nuclease contamination
- Standard 1.1 x 25 x 76 mm size including the PVDF layer
- Corner chamfer (upper right corner) for unambiguous orientation
- High efficiency hydrophobic binding
- Supports all microarray manufacturing methods based on contact and non-contact printing
- Low background when used with Arrayit SpotWare Microarray Scanners
- Uniform feature size due to the homogeneity of the surface
- 25 substrate slides per box
- Anti-static and impact resistant packaging
- Indefinite shelf life when stored dark and dry at room temperature
Technical Information
Arrayit SuperPVDF slides are compatible with most microarray scanners including the InnoScan Microarray Scanners, SpotLight Microarray Scanner and the SpotWare Colorimetric scanner. Fluorescence, colorimetric and chemiluminescence detection methods can be used with this surface. Arrayit SuperPVDF Microarray Substrate Slides are ready to use “right out of the box” without any additional processing or surface preparation. This surface is fully compatible with a variety of manufacturing technologies including Arrayit contact printing as well as non-contact printing such as ink-jetting.
Microarray Printing:
Proteins should be printed in Protein Printing Buffer. If the protein is already in a 1X PBS solution, add an equal volume of 2X PPB printing buffer to achieve a final protein concentration of 0.25-0.5 µg/µl. Arrayit SuperPVDF contains a 150-micron thick layer of polyvinylidene fluoride membrane that has a high binding capacity. When using a robotic microarray printer such as the NanoPrint 2, SpotBot 4 or other microarrayer in conjunction with ArrayIt printing technology, adjust the Z-axis speed to 2 cm/sec or less to avoid denting the membrane surface. Because of the hydrophobic nature of this substrate slide, printing dwell times are flexible. A relative humidity setting of 45-65% RH and ambient temperature of 18-20°C should be used. To maintain protein sample integrity, Arrayit offers protein edition (PRO) NanoPrint 2 and SpotBot 4 microarray printers with 384-well microplate cooling.
Printed Microarray Storage:
Printed microarrays should be incubated overnight at low humidity to drive the hydrophobic binding of printed proteins to the PVDF surface. Refrigeration can be used, but Arrayit printing buffers are designed to stabilize proteins and DNA for long-term storage at room temperature. Vacuum packing can also be used; make sure microarrays are dry prior to vacuum packing. Store printed microarrays unblocked and un-washed, performing blocking and washing steps just prior to incubation or hybridization.
SuperPVDF Blocking, Binding and Washing:
Prior to performing binding reactions, protein microarrays printed on SuperPVDF microarray substrate slides should be blocked with SuperProtein blocking buffer for 1 hour and then washed 3 times in 1X Arrayit Protein Microarray Wash Buffer (Cat. PMWB) for 5 minutes using a high throughput wash station. Make sure to keep the PVDF surface hydrated at all times during the blocking, binding and washing reactions. SuperPVDF should only be allowed to dry after printing and prior to scanning.
Multiplexing Reaction Tools:
SuperPVDF slides are compatible with the multiwell reaction tools AHC1X16, AHC1X24, AHC4X24, and MMH4X24. Using these tools and the right microarray printer programming makes it possible to run up to 24 individual microarray specimens on a single slide and up to 96 individual microarray specimens in a single Arrayit hybridization cassette.
Example Protocol:
See the schematic in Figure 1 as an example of a PVDF assay to detect antibodies that bind to target antigens printed on the SuperPVDF slide.
Figure 1. Schematic illustration of a PVDF antibody binding assay. Antigens are printed and attached to the SuperPVDF surface. Patient serum containing antibodies is incubated with the printed microarray. After the primary antibody binding reaction, a secondary antibody containing a detectable linked conjugate is incubated and subsequently developed to allow fluorescent or colorimetric detection of the antibody. The use of an internal calibration curve for each subclass of antigen/antibody reactions on the microarray provides a unique advantage over traditional immunoassays, where the standard curve and the sample are processed in separate tubes or in separate wells.
Figure 2. Correct substrate slide orientation. Shown is a graphic of two Arrayit 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 slides is suitable for printing. Please print on the correct side only.
Figure 3. Arrayit SuperPVDF Microarray Substrate Slides (1 x 25 x 76 mm) contain a 150 µm layer of polyvinylidene fluoride (PVDF) membrane laminated on the top surface of the glass slide. The corner chamfer can be used to assist in proper side orientation. Microarray printing should be performed on the PVDF side of the slide only.
Short Protocol:
- Print biomolecules into a microarray using a SpotBot 4 Protein Edition Microarrayer, NanoPrint 2 or other suitable microarray printer.
- Wash using PBS and Block using SuperProtein Blocking Buffer.
- Perform binding reaction.
- Wash with Protein Microarray Wash Buffer.
- Perform reaction with secondary labeled conjugated antibody.
- Wash.
- React detection reagents (colorimetric, fluorescent, chemiluminescent)
- Scan with SpotWare, SpotLight or Innoscan series microarray scanners (or other suitable slide based microarray scanner) and analyze the microarray data.
Complete Protocol:
1. Print purified proteins suspended in Protein Printing Buffer (Cat. PPB) at 0.1-1.0 µg/µl as a final concentration from an MMP384 microplate with no more than 15 µl per well with a SpotBot 4 Protein Edition Microarrayer, NanoPrint 2 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 1,011 proteins per mm2 of substrate. 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 printed microarrays by adding 400 ml of 1X Protein Microarray Wash Buffer (Cat. PMWB) in a High Throughput Wash Station and wash with gentle agitation for 5 minutes. Remove printed microarray slides and spin dry for 5 seconds using a Microarray Centrifuge. Use 4 µl of SuperProtein blocking buffer per square centimeter of coverslip and incubate for 1 hour and then wash for 5 minutes with Protein Microarray Wash Buffer (Cat. PMWB) 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 containing 10 ml of blocking buffer.
4. Decant the reaction mixtures and wash the slides in Protein Microarray Wash Buffer (Cat. PMWB) 3 times for 5 minutes per wash in a High Throughput Wash Station.
5. React the microarray with an enzyme- or fluorescence-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 such as the TrayMix. A 60-minute incubation at 25-37°C is usually sufficient to obtain a strong signal.
6. Wash away the secondary antibody mixture using a High Throughput Wash Station and Protein Microarray Wash Buffer (Cat. PMWB) 3 times for 5 minutes per wash.
7. Use the appropriate enzymatic developer for colorimetric detection. Fluorescent assays can be scanned directly. For quantitative work, pre-determine the development time by conducting a pilot study. It is important that all microarrays to be compared are developed for the same amount of time. It is not always necessary to see signal by eye because the SpotWare Colorimetric Microarray Scanner can detect signals that are invisible to the naked eye.
8. To prevent excessive color development, immerse the microarrays in distilled water 2 minutes. Spin the slides dry in an Arrayit Microarray Centrifuge. Colorimetric microarrays can be scanned using a SpotWare Colorimetric Microarray Scanner. Fluorescent based detection systems use a SpotLight or Innoscan series microarray scanner.
Arrayit SuperNylon Microarray Substrate Slides offer a highly innovative membrane surface for researchers wishing to utilize the time-tested attributes of positively charged nylon membranes in a microarray format. Arrayit SuperNylon can be used to attach DNA, protein, carbohydrate and any other molecule that has been shown to bind nylon in traditional filter assays. The high binding capacity of the 150 µm nylon layer is complimented by a polished atomically smooth glass substrate slide for optimal performance compatibility with all standard glass substrate slide microarrayers and scanners.