Arrayit SuperAvidin Blocking Buffer is an advanced protein and small molecule blocking buffer that blocks and inactivates Superavidin microarray substrate slides prior to staining, greatly reduces background for all applications, 250 ml of 1X buffer solution.
Reagents - Microarray Buffers - Blocking Solutions - BlockIt™ and BlockIt™ HT Blocking Buffers for DNA and Protein Microarrays
Arrayit has developed and manufactured an advanced microarray blocking solution designed to inactivate reactive groups remaining post-printing on SuperAvidin Microarray Substrate Slides and other glass microarray substrate slide surfaces containing biotin-binding proteins. SuperAvidin Blocking Buffer greatly reduces background noise while maintaining full signal intensities for a wide spectrum of DNA and protein microarray applications. 250 ml of 1X buffer.
Table of Contents
- Introduction
- Quality Control
- Product Description
- Technical Assistance
- Short Protocol
- Complete Protocol
- Short Protocol DNA
- Complete Protocol DNA
- Troubleshooting Tips
- Recommended Requirements
- Ordering Information
- Warranty
Introduction
Congratulations on taking a big step towards improving the economies of scale, quality and speed of your genomics research. This booklet contains a complete set of protocols outlining the steps and principles needed to use the Arrayit brand BlockIt blocking buffer for microarray surface chemistries. This product is design for use prior to incubation and hybridization reactions involving protein and DNA microarrays.
Quality Control
Arrayit assures the performance of these products, the finest scientific research went into their development. Store at 4°C, the product has a 1-year shelf life when handled and stored properly.
Product Description
Arrayit brand BlockIt™ buffer is designed for blocking the portions of microarrays that are still reactive after printing to eliminate fluorescent background following the incubation or hybridization reaction of protein and DNA microarrays. Users will appreciate the following features:
- Ultra-pure reagents used for buffer formulation
- Purified by sterile filtration
- Provided as 1X ready to use concentrate
- Recommended for use on all types of protein or DNA microarray glass microarray surface chemistries
- Easy cover slip blocking procedure for minimal reagent consumption
- 250 ml volume provided efficiently blocks thousands of microarrays
- Will not cross react with proteins and DNA
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 email: arrayit@arrayit.com, Monday–Friday PST 8:00am - 8:00pm. Please remember that we want to hear about your successes!
Short Protocol for Protein Microarrays
Wash microarray and then incubate microarray in 1X BlockIt buffer under a coverslip for 1 hour prior to protein binding or hybridization reactions.
Complete Protocol for Protein Microarrays
BlockIt will couple to un-reacted binding groups on microarray surfaces and prevent background fluorescence. Once the printing process is complete, and samples are coupled to the surface, block a microarray surface by using a 1-24 hour incubation at room temperature in 1X BlockIt buffer. Perform this reaction under a coverslip using 4 ul of BlockIt for every cm2 of coverslip area. A Hybridization Cassette should be used to keep the blocking reaction from drying out. After blocking, wash the microarrays to remove excess buffer. Wash 2 times for 15 minutes each in a High Throughput Wash Station in 1X PBST to remove unbound protein molecules and buffer components from the microarray. Protein and DNA binding to the SuperEpoxy 3 surface is extremely stable and the microarrays can be washed, blocked and reacted without sufficient loss of coupled protein.
NOTE: BlockIt can be used as a reaction buffer for all protein microarray applications.
Figure 1. BlockIt™ background fluorescence reduction on SuperAmine (left panel) and SuperEpoxy (right panel) microarray glass substrate slides. Blocking reactions were performed at room temperature for 16 hours. 10 µl of BlockIt™ was used under 22 x 22 mm coverslips. The substrate slides were washed in 1x PBS in a High Throughput Wash Station for 10 minutes at medium speed on a stir plate. The substrate was spun in a Microarray High-Speed Centrifuge until dry. A total of 30 µl of 0.6% Cy3-labeled IgG was reacted across the substrate slides under 20 x 60 mm coverslips. The IgG was suspended in a 1% BSA in PBS solution. The solution was allowed to react for 4 hours at room temperature protected from light. The substrate slides were washed in 1x PBS in a High Throughput Wash Station for 10 minutes and spun dry. The substrate slides were scanned using a PerkinElmer ScanArray Express set at 70% PMT and 90% laser power at a 10 µm resolution. The signal intensities (see Table) were averaged across a circular area with an 8,575 µm diameter. The upper panel shows the scanned images and the table records the signal intensities. BlockIt™ blocking solution reduces background noise by 2-10 fold depending on the surface.
Glass Substrate Slide |
- BlockIt™ (fluorescent counts) |
+ BlockIt™ (fluorescent counts) |
SuperAmine Microarray Substrates |
5,586 |
2,924 |
SuperEpoxy Microarray Substrates |
2,079 |
698 |
Short Protocol for DNA Microarrays
Incubate microarray in 1X BlockIt™ buffer under a coverslip or in a microarray reaction tray for 15-60 min prior to hybridization.
Complete Protocol for DNA Microarrays
BlockIt will couple to the un-reacted groups on the surfaces and prevent background fluorescence. Once the printing process is complete, and samples are bound to the surface use a 1-24 hour incubation at room temperature of 1X BlockIt buffer. Perform this reaction under a cover slip using 4 ul of BlockIt for every cm2 of coverslip area. A Hybridization Cassette can be used to keep the blocking reaction from drying out. After blocking wash the printed microarrays in a High Throughput Wash Station using the protocol recommended by the manufacturer. DNA binding to the SuperAmine 3, SuperAldehyde 3 and SuperEpoxy 3 surfaces is extremely stable and the microarrays can be washed, blocked and reacted without significant loss of coupled DNA. After blocking, wash and process the microarrays according to the manufacturers recommendation to remove excess buffer and unbound sample.
Figure 2. Shown is a microarray blocking reaction using BlockIt™ Microarray Blocking Buffer. Three DNA microarrays are incubated overnight with gentle agitation in a square culture dish containing 30 ml of 1X BlockIt™ to inactivate the microarray surface and reduce fluorescent background during the hybridization reaction. BlockIt™ molecules bind covalently to un-reacted groups on the microarray surface, preventing non-specific binding of probe molecules during hybridization. Unlike other blocking solutions that contain cloudy additives and impure reagents, BlockIt™ is perfectly clear solution of highly purified and soluble components that provides a vastly superior blocking function. BlockIt™ is an all-purpose blocking solution for DNA, protein, and other types of microarrays.
Troubleshooting Tips
If microarray shows high background:
- Confirm that the intrinsic background of the microarray is low to start with.
- Be sure to not let any sample dry on the microarray surface during the binding reaction.
- Make sure to purify away unincorporated dyes in the labeling reaction.
- Take care to block for the recommended time (1 hour).
- Pay attention to the proper handling and shelf life of the buffer
(1 year shelf life, Store at 4 C).
- Do not skip washing the microarray after the blocking step.
Recommended Products:
Reagents - Microarray Buffers - Blocking Solutions - BlockIt™ Plus Microarray Blocking Buffer for Enhanced DNA and Protein Glass Substrate Slide Microarrays
BlockIt™ Plus microarray blocking solution designed to inactivate reactive groups remaining post-printing on SuperEpoxy 3, SuperAmine 3, SuperAldehyde 3, SuperNitro, SuperPVDF and SuperNylon and other microarray substrate slide surfaces. BlockIt™ Plus Blocking Solution greatly reduces background noise while maintaining full signal intensities for a wide spectrum of microarray applications. 250 ml of 1X buffer.
Table of Contents
- Introduction
- Quality Control
- Product Description
- Technical Assistance
- Short Protocol
- Complete Protocol
- Short Protocol DNA
- Complete Protocol DNA
- Troubleshooting Tips
- Recommended Requirements
- Ordering Information
- Warranty
Introduction
Congratulations on taking a big step towards improving the economies of scale, quality and speed of your microarray research. This booklet contains a complete set of protocols outlining the steps and principles needed to use BlockIt™ Plus blocking buffer for microarray surface chemistries. This product is design for use prior to incubation and hybridization reactions involving protein and DNA microarrays.
Quality Control
Arrayit assures the performance of these products, the finest scientific research went into their development. Store at 4°C, the product has a 1-year shelf life when handled and stored properly.
Product Description
Arrayit BlockIt™ Plus blocking buffer is designed for blocking the portions of microarrays that are still reactive after printing to eliminate fluorescent background following the incubation or hybridization reaction of protein and DNA microarrays. Users will appreciate the following features:
- Often works when regular BlockIt™ is not sufficient
- Ultra-pure reagents used for buffer formulation
- Provided as 1X ready to use concentrate
- Recommended for use on all types of protein or DNA microarray glass microarray surface chemistries
- Easy cover slip or petri dish blocking procedure for minimal reagent consumption
- 250 ml volume provided can efficiently block thousands of microarrays
- Will not cross react with proteins and DNA
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 email: arrayit@arrayit.com, Monday–Friday PST 8:00am - 8:00pm. Please remember that we want to hear about your successes!
Short Protocol for Protein Microarrays
Wash microarray and then incubate microarray in 1X BlockIt™ Plus buffer under a coverslip for 1 hour prior to protein binding or hybridization reactions.
Complete Protocol for Protein Microarrays
BlockIt will couple to un-reacted binding groups on microarray surfaces and prevent background fluorescence. Once the printing process is complete, and samples are coupled to the surface, block a microarray surface by using a 1-24 hour incubation at room temperature to 4 degress celcius in 1X BlockIt™ Plus buffer. Perform this reaction under a coverslip using 4 ul of BlockIt for every cm2 of coverslip area, also compatible with LifterSlips. A Hybridization Cassette can be used to keep the blocking reaction from drying out. After blocking, wash the microarrays to remove excess buffer. Wash 2 times for 15 minutes each in a High Throughput Wash Station in Protein Microarray Wash Buffer to remove unbound protein molecules and buffer components from the microarray. Protein and DNA binding to the SuperEpoxy 3, SuperNitro and SuperPVDF surfaces is extremely stable and the microarrays can be washed, blocked and reacted without sufficient loss of coupled protein.
NOTE: BlockIt™ Plus is not recommended to be used as a reaction buffer for protein microarray applications.
Short Protocol for DNA Microarrays
Incubate microarray in 1X BlockIt™ Plus buffer under a coverslip or in a microarray reaction tray for 15-60 min prior to hybridization.
Complete Protocol for DNA Microarrays
BlockIt™ Plus will couple to the un-reacted groups on the surfaces and prevent background fluorescence. Once the printing process is complete, and samples are bound to the surface use a 1-24 hour incubation at room temperature of 1X BlockIt Plus buffer. Perform this reaction under a cover slip using 4 ul of BlockIt for every cm2 of coverslip area. A Hybridization Cassette can be used to keep the blocking reaction from drying out. After blocking wash the printed microarrays in a High Throughput Wash Station using the protocol recommended by the manufacturer. For best results use Arrayit DNA Microarray Wash Buffers. DNA binding to the SuperAmine 3, SuperAldehyde 3 and SuperEpoxy 3 surface is extremely stable and the microarrays can be washed, blocked and reacted without significant loss of coupled DNA. After blocking, wash and process the microarrays according to the manufacturers recommendation to remove excess buffer and unbound sample.
Figure 1. Shown is a microarray blocking reaction using BlockIt™ Plus Microarray Blocking Buffer. Three DNA microarrays can be incubated overnight with gentle agitation in a square culture dish containing 30 ml of 1X BlockIt™ to inactivate the microarray surface and reduce fluorescent background during the hybridization reaction. BlockIt™ Plus molecules bind covalently to un-reacted groups on the microarray surface, preventing non-specific binding of probe molecules during hybridization. BlockIt™ Plus is not a clear solution, but contains highly purified and soluble components that provide a highly efficient blocking function. BlockIt™ Plus is an all-purpose blocking solution for DNA, protein, and other types of microarrays and is used when blocking with BlockIt™ shows to be inefficient.
Troubleshooting Tips
If microarray shows high background:
- Confirm that the intrinsic background of the microarray is low to start with.
- Be sure to not let any sample dry on the microarray surface during the binding reaction.
- Make sure to purify away unincorporated dyes and other conjugates such as biotin, AP and HRP in the labeling reaction.
- Take care to block for the recommended time (at least 1 hour).
- Pay attention to the proper handling and shelf life of the buffer
(1 year shelf life, Store at 4 C).
- Do not skip washing the microarray after the blocking step.
Recommended Products:
Reagents - Microarray Buffers - Blocking Solutions - SuperProtein Blocking Buffer to Enhance Signal Strength and Reduce Non-Specific Binding on Nitrocellulose Microarrays
Designed to block SuperProtein, SuperPVDF, SuperNitro membrane based microarray surface chemistry prior to protein binding reactions for ultra low colorimetric background. For glass based protein microarrays use BlockIt. For assays sensitive to low amounts of detergent use SuperProtein Blocking Buffer.
Table of Contents
- Introduction
- Quality Control
- Product Description
- Technical Assistance
- Short Protocol
- Complete Protocol
- Troubleshooting Tips
- Recommended Requirements
- Ordering Information
- Warranty
Introduction
Congratulations on taking a big step towards improving the economies of scale, quality and speed of your proteomics research. This booklet contains a complete set of protocols outlining the steps and principles needed to use the Arrayit brand blocking buffer for membrane-based surface chemistry. This product is design for use prior to incubation and hybridization reactions involving protein and DNA microarrays.
Quality Control
Arrayit assures the performance of these products, the finest scientific research went into their development. Store at 4 Degrees Celsius, the product has a 1-year shelf life when handled and stored properly.
Product Description
The buffer is designed for blocking the portions of microarrays that can bind proteins after printing to eliminate background following the incubation or hybridization reaction of protein microarrays. Users will appreciate the following features:
- Ultra-pure reagents used for buffer formulation
- Purified by sterile filtration
- Provided as 1X ready to use concentrate
- Recommended for use on SuperProtein Substrates, but compatible with other protein membrane based microarray surface chemistries
- Easy coverslip blocking procedure
- 250 ml volume provided efficiently blocks thousands of microarrays
- Will not react with proteins and DNA
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 email: arrayit@arrayit.com, Monday–Friday PST 8:00am - 8:00pm. Please remember that we want to hear about your successes!
Short Protocol for Protein Microarrays
Wash microarray with PBS and then incubate microarray in 1X blocking buffer under a coverslip for 1 hour prior to protein binding reactions.
Complete Protocol for Protein Microarrays
Protein binding to our membrane surfaces is stable and the microarrays can be washed, blocked and reacted without sufficient loss of coupled protein. Block the printed microarray surface using a 1-hour incubation at room temperature in 1X SuperProtein Blocking buffer. Perform this reaction under a coverslip using 4 ul of buffer for every cm2 of coverslip area. Once the printing process is complete, wash the printed microarrays 2 times for 15 minutes each in a High Throughput Wash Station in 1X PBS to remove unbound protein molecules and buffer components from the SuperProtein Substrates.A Hybridization Cassette can be used to keep the blocking reaction from drying out. SuperProtein blocking buffer will couple to un-reacted binding sites of the surface and prevent unspecific binding and background. After blocking, wash the microarrays to remove excess buffer. Washing three times for 1 min each at room temperature with 1X PBS in a High Throughput Wash Station. DO NOT USE DETERGENT BASED buffers on the SuperProtein nor SuperPVDF, low concentrations are compatible with SuperNitro.
Troubleshooting Tips
If microarray shows high background:
- Confirm that the intrinsic background of the microarray is low to start with.
- Be sure to not let any sample dry on the microarray surface during the binding reaction.
- Make sure to purify away unincorporated components of any labeling reactions.
- Take care to block for the recommended time (1 hour).
- Pay attention to the proper handling and shelf life of the buffer (1 year shelf life, Store at 4 C).
- Do not skip washing the microarray after the blocking step.
- If colorimetric labeling was used, be sure to optimize development times
Recommended Products
Ordering Information
|
Product
|
Description
|
Catalog
ID
|
|
BlockIt™ Blocking Solution
|
Arrayit BlockIt™ Blocking
Solution is an advanced microarray blocking solution designed to inactivate
reactive groups remaining post-printing on SuperEpoxy, SuperAmine, SuperAldehyde
and other glass microarray substrate slide surfaces. BlockIt™ Blocking
Solution greatly reduces background noise while maintaining full signal
intensities for a wide spectrum of DNA and protein microarray applications.
Ultra-high purity blocking buffer for coverslip and solution-based blocking
of protein and DNA microarrays, 250 ml of 1X buffer, ships on cold packs.
|
BKT
|
|
BlockIt™ HT Blocking
Solution
|
Arrayit BlockIt™ HT
Blocking Solution is an advanced microarray blocking solution designed to inactivate
reactive groups remaining post-printing on SuperEpoxy, SuperAmine,
SuperAldehyde and other glass microarray substrate slide surfaces. BlockIt™
Blocking Solution greatly reduces background noise while maintaining full
signal intensities for a wide spectrum of DNA and protein microarray
applications. High purity blocking buffer for bulk solution high-throughput
(HT) blocking of protein and DNA microarrays for use with the High Throughput
Wash Station, 1000 ml of 1X buffer, ships on cold packs.
|
BKTHT
|
|
BlockIt™ Plus Blocking
Buffer
|
Arrayit BlockIt™ Plus
Blocking Solution often works when BlockIt™ is shown to be less efficient
than required. Greatly reduces background noise while maintaining full signal
intensities for a wide spectrum of DNA and protein microarray applications.
Ultra-high purity blocking buffer (translucent), for blocking of protein and
DNA microarrays, 250 ml of 1X buffer, ships on cold packs.
|
BKTP
|
|
Microarray Blocking Buffer
Plus Automation Formulation
|
Arrayit Microarray
BlockIt™ Plus Blocking Solution Automation Formulation should be used in
cases where a greatly reduced background is required, while maintaining full
signal intensities for a wide spectrum of DNA and protein microarray
applications particularly those utilizing workstations, liquid handling
robots, multiplex hybridization cassettes and other high-throughput hardware
and automation equipment. 1L of 1X buffer arrives pre-mixed and ready to use,
ships on cold packs.
|
BKTPL
|
|
SuperProtein Substrate
Blocking Buffer
|
Arrayit SuperProtein
Substrate Blocking Buffer is an advanced microarray blocking solution
designed to inactivate reactive groups remaining post-printing on our
membrane-based microarray surfaces such as nitrocellulose, PVDF and nylon.
SuperProtein Blocking Solution greatly reduces background noise while
maintaining full signal intensities for a wide spectrum of protein microarray
applications. 250 ml of 1X buffer, ships on cold packs.
|
SPBB
|
|
SuperStreptavidin
Microarray Blocking Buffer
|
Arrayit SuperStreptavidin Microarray
Blocking Buffer is an advanced protein and small molecule blocking buffer
that blocks and inactivates SuperStreptavidin microarray substrate slides
prior to staining, greatly reduces background for all applications, 250 ml of
1X buffer solution, ships on cold packs.
|
SBB
|
|
SuperAvidin Blocking
Buffer
|
Arrayit SuperAvidin
Blocking Buffer is an advanced protein and small molecule blocking buffer
that blocks and inactivates SuperAvidin microarray substrate slides prior to
staining, greatly reduces background for all applications, 250 ml of 1X
buffer solution, ships on cold packs.
|
ABB
|
|
ChemBlock™
Microarray Chemical Blocking Buffer
|
Arrayit ChemBlock™
Microarray Blocking Buffer features a proprietary mixture of chemical
blocking agents, buffering components and detergents to inactive aldehyde and
epoxide reactive groups after microarray printing and before DNA and protein
microarray reactions to eliminate background binding observed with
traditional protein blocking buffers. ChemBlock™ is highly recommended for
serum reactions and other protein microarray applications requiring uniform
and extremely low background signals across a large number of blood
specimens, 0.2 µm filtered, store at -20°C, 1 liter of 1X solution, ships on
cold packs.
|
CHE
|
Warranty
Arrayit life sciences
products are sold for research purposes only. Arrayit brand products have been
scientifically developed and are sold for research purposes. Extreme care and
exact attention should be practiced in the use of the materials described herein.
All Arrayit brand products are subject to extensive quality control and are
guaranteed to perform as described when used properly. Any performance issues
should be reported to Arrayit immediately. Arrayit’s 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. Pricing may vary up to 30% due to costs associated
with distribution, import taxes, duties, customs clearance and shipping.