Arrayit Corporation ARYC Protein A Protein G Microarray Substrate Slides - Staphylococcus Aureus Immunoglobulin IgG Antibodies Binding Surface


	Arrayit (ARYC) leads and empowers the genetic, research, pharmaceutical, and diagnostic communities through the discovery, development and manufacture of proprietary life science technologies and consumables for disease prevention, treatment and cure.

		- Powerful Science for Life™ Share \|

Products - Microarray Substrates & Slides - Protein A/G Substrates

ArrayIt® Protein A/G Microarray Substrates contain a highly uniform mono layer of Recombinant Protein A/G over the surface of the substrate for microarray applications. The surface is designed to bind human immunoglobulin proteins IgG₁, IgG_2,IgG_3,IgG₄and all types of total IgG. The open platform microscope slide size format format makes this product compatible with all standard microarrayer printers, scanners and other microscope slide processing and detection equipment.

Table of Contents

Introduction
Quality Control
Product Description
Protocol Protein Microarray
Technical Specifications
Technical Assistance
Recommended Equipment
Ordering Information
Storage Conditions
Warranty

Introduction
Congratulations on taking a big step towards improving the affordability, quality and speed of your genomics, biomedical, pharmaceutical and agricultural research. This booklet contains all the information required to take full advantage of ArrayIt^® Protein A/G Microarray Substrates.

Quality Control
Arrayit takes every measure to assure the quality of our ArrayIt^® Brand Super Microarray Substrates. The finest microarray biochip cleanroom research was used to develop these products. Rigorous quality control monitoring on a substrate-by-substrate basis guarantees that each these products conforms to the highest industry standards.

Product Description
This surface starts with ArrayIt SuperClean glass then it is activated with Protein A/G. Gene fusion of the Fc-binding domains of Protein A and Protein G has resulted in production of a structurally and functionally chimeric protein with broader binding than either Protein A or Protein G alone. During fusion, the Protein G gene sequence coding for the serum albumin-binding site is eliminated. This protein is expressed in E.Coli and secreted into the surrounding medium during fermentation. The product obtained is consistent in quality and yield because the bacterial host is engineered to be deficient in major proteolytic activities. Binding is less pH-dependent than either Protein A or Protein G alone, occurring well at pH 5-8. The extended Fc-binding properties of Protein A/G make it a popular tool in the investigation and purification of immunoglobulins. Protein A/G binds to all human IgG subclasses, IgA, IgE, IgM and to a lesser extent IgD; however, it does not bind mouse IgA, IgM or murine serum albumin. Protein A/G is an excellent tool for purification and detection of mouse monclonal antibodies from IgG subclasses without interference from these other serum proteins. Individual subclasses of mouse monoclonals are most likely to have stronger affinity to this chimeric protein than to either Protein A or Protein G. The Protein A/G on the substrate is covalently bound to the surface and will not flake, crack or peel off the slide. SuperClean substrate as the foundation for this product is a highly polished glass surface with an optical smoothness tolerance of 50 angstroms over the entire 25 mm x 76 mm surface area for optimum homogeneity; this new surface sets the highest quality standard for Protein A/G-activated substrates for microarray applications. These substrates are packed in white slide boxes, vacuum packed and should be stored at 4 C.

Short Protocol Protein (Steps 1-7)
1. Suspend protein samples in Protein Printing Buffer at 0.25-1 µg/µl.
2. Print protein samples onto Protein A/G Substrates.
3. Block with BlockIt buffer and then wash and dry the printed microarrays.
4. React the processed microarrays with fluorescent samples.
5. Wash the microarrays to remove un-reacted fluorescent material.
6. Scan the microarray to produce a fluorescent image.
7. Quantitate and model the fluorescent data.

Complete Protocol Protein (Steps 1-7)
1. Suspend protein samples to be spotted onto the Protein A/G substrate in 2X Protein Printing Buffer for a final protein concentration of 0.25-1 µg/µl. Perform transfers careful to prevent protein denaturation. Mix samples thoroughly prior to printing and transfer into 384-well microplates at 5-10 µl per well. Keep samples cold (4°C) at all times during sample preparation.

2. Print protein samples onto Protein A/G Substrates. Use a Protein Edition Microarrayer such as the SpotBot® Pro or NanoPrint Protein Edition to print the protein samples on the Protein A/G Substrates. Place the 384-well microplates onto the platen of the instrument and print the samples onto the glass Protein A/G Substrate slides. For best results, make sure the microplates remain at 4°C for the duration of the print run. Following the print run, incubate the printed substrate slides on the robot platen for 1 hour at ambient humidity, to allow binding to take place.

3.Activate the Microarrays by mixing for 60 min in Protein Microarray Activation Buffer (Cat. PMAB). The activation step can be performed under a cover slip or in batch mode using 450 ml of Protein Microarray Activation Buffer with gentle mixing in a High-Throughput Wash Station. Following activation, wash the substrates two times for 5 min in Protein Microarray Wash Buffer (Cat. PMWB) and one time for 1 sec in Protein Microarray Rinse Buffer (Cat. PMRB). All washes can be performed using 25 ml volumes in a petri dish or 450 ml volumes in a High-Throughput Wash Station (Cat. HTW ) with gentle agitation. After washing, spin for 1 sec in a Microarray High-Speed Centrifuge to dry the substrates.

4. React the activated microarrays with sample of choice. Samples can be prepared by mixing 5-25 µg of labeled protein with Protein Microarray Reaction Buffer (Cat. PMRB), making sure not to dilute the Protein Microarray Reaction Buffer by more than 1.5-fold. The same volume of protein sample should be used for all reactions in which comparative information is sought. Reactions can be performed using LifterSlip™ (elevated cover slips) or multi-well reaction cassettes. Binding reactions may also be performed using large sample droplets or a variety of custom gaskets. Undiluted or diluted samples can be used depending on the sample concentration and protein expression level. Typically 5 µg of labeled protein is sufficient to obtain strong signals. Binding reactions should be allowed to proceed for 1 hour to overnight . Following the reaction step, wash the microarrays three times for 3 min in Protein Microarray Wash Buffer (Cat. PMWB) with gentle agitation. Washes can be performed using 25 ml volumes in a petri dish or 450 ml volumes in a High-Throughput Wash Station with gentle agitation. After washing, spin briefly in a Microarray High-Speed Centrifuge to dry.

5. Detection ith a fluorescent streptavidin secondary reagent: Prepare the secondary antibody reagent by diluting the Cy3™- or Cy5™-streptavidin conjugate to a final concentration of 1 µg/ml in Protein Microarray Reaction Buffer (Cat. PMRB). A one thousand-fold (1:1,000) dilution of the 1 mg/ml Cy3™-Streptavidin staining reagent from Jackson ImmunoResearch (Cat.016-160-084 ) works well for this application. A 1.0 ml volume of secondary reagent is prepared by mixing 1.0 µl Cy3™-streptavidin in 1 ml of Protein Microarray Reaction Buffer. Mix by inverting the tube 10 times. Stain the microarrays using 25-125 µl of diluted secondary reagent. The staining volume will depends on whether a single or multi-well microarray format is being used. Stain for 60 min at room temperature with gentle agitation. Following the staining step, wash the microarrays three times for 1 min in Protein Microarray Wash Buffer (Cat. PMWB) and one time for 1 sec in Protein Microarray Rinse Buffer (Cat. PMNB) with gentle agitation. Washes can be performed using 25 ml volumes in a petri dish or 450 ml volumes in a High-Throughput Wash Station with gentle agitation. After washing, spin briefly in a Microarray High-Speed Centrifuge to dry.

6. Scan the microarrays for fluorescence emission. Scan the Microarrays using an Arrayit SpotLight™or Arrayit InnoScan® microarray scanner set to detect Cy3™ or Cy5™. These slide substrates can also be read with other brands of microarray scanner compatible with the open platform substrate slide 25 x 76 mm format. Scanner settings should be adjusted to minimize saturated signals to 1% or less. Multiple scans can be taken to capture data at different sensitivity levels. All data files should be saved as 16-bit TIFF images for data analysis.

Technical Specifications:
Protein A/G Microarray Substrates are not coated microscope slides! Atomically flat polished glass is used as the starting point for this product. Glass substrates of this quality routinely retail at $50-100 each from leading optical suppliers. Users will benefit from the following technical features:

Glass polished to atomic smoothness (±20 angstroms)
Only polished glass surface in the microarray industry
Polishing improves topology and uniformity – keys to precise data
Homogenous Protein A/G proteins providing binding to human IgG₁_,IgG₂_,IgG₃_,IgG₄
Protein A/G molecules are covalently bound to the glass substrate
Protein A/G monolayer contains 1.1 x 10^10 proteins/mm2
Protein A/G monolayer covers entire 25 x 76 mm surface area
Vastly superior to unpolished optical quality glass from other vendors
Compatible with both contact and non-contact printing
Manufactured in a state-of-the-art class 100 cleanrooms
Ultra-low intrinsic fluorescence and background noise
Open platform dimensions compatible with all major brands of microarrayers, scanners and other microscope slide size processing tools.
Precise physical dimensions (25 ± 0.2 mm x 76 ± 0.3 mm x 0.940 mm ± 0.025 mm)
Proprietary corner chamfer (1.4 mm) provides unambiguous side and end orientation to simplify printing and processing
Finished edges enhance user safety
Glass with excellent refractive index, transmission and hardness specifications
Sophisticated anti-static packaging improves usability
Offered with or without bar-codes
Custom laser and chrome fiducials available upon request
High-volume 100,000 piece per month manufacturing capabilities
Product arrives “ready to use” with no additional processing required

Figure 2. Atomic Force Microscopy (AFM) analysis. Shown are AFM scans of the SuperClean Microarray Substrate by Arrayit ArrayIt Division, Sunnyvale CA. Data are coded to a rainbow intensity scale, such that red data represents 5.0 nm or 50 angstroms. The substrates have an average smoothness of 2.0 nm or 20 angstroms, which corresponds to about 10 silicon dioxide bonds.

Figure 3. 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.

Technical Assistance
Please contact us if you have any comments, suggestions, or if you need technical assistance. By electronic mail: arrayit@arrayit.com(under the subject heading please type "ArrayIt® technical assistance"). By telephone: (408) 744-1331, Monday–Friday PST 9:00am - 4:30pm. Please remember that we want to hear about your successes!

Scientific Publications
Click here and here for recent scientific publications using ArrayIt® brand microarray products from Arrayit International, Inc for protein A/G experimentation.

Ordering Information

Product	Description	Catalog ID	Price (U.S. dollars)
Protein A/G Microarray Substrates(Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, box of 25.	AGMS	$572
Protein A/G Microarray Substrates Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity binding and low non-specific binding over the entire 25 x 76 mm area, box of 25, barcoded.	AGMSBC	$599
Protein A/G Microarray Substrates (Box of 5)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity binding and low non-specific binding over the entire 25 x 76 mm area, 5 substrates per box.	AGMSF	$184
Protein A/G Microarray Substrates Barcoded (Box of 5)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity binding and low non-specific binding over the entire 25 x 76 mm area, box of 5, barcoded.	AGMSFBC	$189
MirrorProtein A/G Microarray Substrates(Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area. Enhanced signals via reflective backside mirror coating. Box of 25.	AGMSMR	$572
MirrorProtein A/G Microarray Substrates Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area. Enhanced signals via reflective backside mirror coating. Box of 25, barcoded.	AGMSMRBC	$599
MirrorProtein A/G Microarray Substrates (Box of 5)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area. Enhanced signals via reflective backside mirror coating. Box of 5.	AGMSMRF	$184
MirrorProtein A/G Microarray Substrates Barcoded (Box of 5)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area. Enhanced signals via reflective backside mirror coating. Box of 5, barcoded.	AGMSMRFBC	$189
Protein A/G Microarray Substrates 4-Well Mask(Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 4-well mask, Box of 25.	AGMSM4	$578
Protein A/G Microarray Substrates 4-Well Mask Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 4-well mask, Box of 25, barcoded.	AGMSM4BC	$604
Protein A/G Microarray Substrates 12-Well Mask (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over a 24 x 60 mm reactive area, 25 x 76 mm, 12-well mask, box of 25.	AGMSM12	$583
Protein A/G Microarray Substrates 12-Well Mask Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 12-well mask, Box of 25, barcoded.	AGMSM12BC	$609
Protein A/G Microarray Substrates 16-Well Mask (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 16-well mask, Box of 25, barcoded.	AGMSM16	$588
Protein A/G Microarray Substrates 24-Well Mask (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 24-well mask, Box of 25.	AGMSM24	$593
Protein A/G Microarray Substrates 48-Well Mask (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 48-well mask, Box of 25.	AGMSM48	$599
Protein A/G Microarray Substrates 48-Well Mask Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 48-well mask, Box of 25, barcoded.	AGMSM48BC	$625
Protein A/G Microarray Substrates 64-Well Mask(Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 64-well mask, Box of 25.	AGMSM64	$604
Protein A/G Microarray Substrates 192-Well Mask (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 192-well mask, Box of 25.	AGMSM192	$609
Protein A/G Microarray Substrates 192-Well Mask Barcoded (Box of 25)	A polished atomically flat glass with a covalently bound Protein A/G layer for high IgG binding capacity and low non-specific binding over the entire 25 x 76 mm area, 192-well mask, Box of 25, barcoded.	AGMSM192BC	$635
Protein Microarray Activation Buffer	Protein Microarray Activation Buffer for activating microarray surfaces prior to use,250 ml of 1X solution, 0.1 µm filtered, arrives ready to use, store at 4°C.	PMAB	$294
Protein Microarray Wash Buffer	Protein Microarray Wash Buffer for washing after activation and reaction steps, 1000 ml of 1X solution, 0.1 µm filtered, arrives ready to use, store at 20-25°C.	PMWB	$100
Protein Microarray Reaction Buffer	Protein Microarray Reaction Buffer for reacting protein samples and secondary staining reagents, 250 ml of 1X solution, 0.1 µm filtered, arrives ready to use, store at 20-25°C.	PMRB	$89
Protein Microarray Rinse Buffer	Protein Microarray Rinse Buffer for rinsing microarrays prior to scanning, 500 ml of 1X solution, 0.1 µm filtered, arrives ready to use, store at 20-25°C.	PMNB	$74
Substrates Discounts 4-19 boxes, 5% discount off list price 20-39 boxes, 10% discount off list price 40-199 boxes, 15% discount off list price Protein Microarray Buffer Discounts 2-5 bottles, 5% discount off list price 6-10 bottles, 10% discount off list price 11-20 bottles, 15% discount off list price 20+ bottles, 20% discount off list price

*To order ArrayIt® Brand Products: call 408-744-1331, fax 408-744-1711, email arrayit@arrayit.com, or click on the purchase buttons to proceed directly to the shop site.

*International pricing may vary as much as 30% due to import duties, stocking fees and technical support.

*Plus shipping and handling.

Storage Conditions
This surface is manufactured and packaged in state-of-the-art class 100-cleanroom facilities. Store at 4°C and use within 2 months from the date of receipt. Store in the sealed plastic shipping envelope and avoid exposure to elevated temperatures and dust.

Warranty
Arrayit's Super Microarray Substrate products have been scientifically developed and manufactured in state-of-the-art class 100 cleanrooms. All of these products are sold for research purposes. Extreme care and exact attention should be practiced in the use of the materials described herein. All Arrayit products are subject to extensive quality control and are guaranteed to perform as described when used properly. Any problems with these products should be reported to Arrayit immediately. Our liability is limited to the replacement of the product, or a full refund. Any misuse of this product is the full responsibility of the user, and Arrayit makes no warranty or guarantee under such circumstances.