Protein Crystallization

Protein crystallization is a key assay for structural studies of proteins. The protocols for crystallization of protein are challenging due to the stringent requirement for pure samples and control of environmental conditions during the crystallization process. Vapor diffusion using hanging drop is a preferred method for obtaining quality crystals with…

more

ARRAYCAM® MULTIPLEX MICROARRAY IMAGING SYSTEM

Affordable bench-top microarray imager and reagent platform for protein and nucleic acid detection of microarrays printed onto ONCYTE® nitrocellulose coated chips.

more

Protein Microarray Substrates

more

Grace Bio-Labs microarray surface chemistry is based on the well-known protein-binding properties of nitrocellulose. A range of different formulations have…

APPLICATIONS:

Antigen-Capture AssayAntibody Capture AssayRPPA- Reverse Phase Protein MicroarrayLaser micro-dissection RRPAEpitope-mappingBiomarker Discovery and ValidationImmunogen Discovery

DNA Microarray Substrates

more

Epoxy Microarray Slides provide a uniform substrate for a variety of DNA/RNA-based diagnostic applications.

APPLICATIONS:

DNA/Oligonucleotide Microarray ;  microRNA Microarray ;  Single Nucleotide Polymorphism (SNP) Analysis ;  Gene Expression Profiling; 

Microarray Reagents

more

Grace Bio-Labs microarray regents have been specifically formulated to achieve the full potential of porous nitrocellulose, accelerating experimental design and…

APPLICATIONS:

Antigen-Capture Assay Antibody Capture Assay RPPA- Reverse Phase Protein Microarray Laser micro-dissection RRPA Epitope-mapping Biomarker Discovery and Validation Immunogen Discovery

APPLICATIONS:

Antibody capture assay ;  Epitope-mapping ;  Biomarker Discovery and Validation ;  Immunogen Discovery ;  Quantitative multiplex immunoassays ;  Peptide Microarrays ;  Autoantibody profiling ;  Multiplex serological assays; 

ProPlate®

more

ProPlates® were specifically designed to enable automated robotic liquid handling. Two main configurations are available: The ProPlate® Microtiter Plate is comprised…

APPLICATIONS:

ProteomicsProtein MicroarraysProtein expression analysisAntibody profiling cDNA and oligonucleotide arrays

ARRAYCAM® MULTIPLEX MICROARRAY IMAGING SYSTEM

more

Affordable bench-top microarray imager and reagent platform for protein and nucleic acid detection of microarrays printed onto ONCYTE® nitrocellulose coated chips.

APPLICATIONS:

Whole Proteome Screening;  Vaccine Development;  RPPA: Reverse Phase Protein Array;  Biomarker Discovery and Validation;  Epitope Mapping; 

NanoParticle Fluorescent Calibration Slide

more

Photostable nanoparticles arrayed on glass slides for calibration of fluorescence imaging systems and quantitative analysis.

APPLICATIONS:

Calibration of Microarray Scanners ;  Quantitative Microarray Analyses Microscope Focal Plane Adjustment;  Microscope Focal Plane Adjustment; 

CoverWell Perfusion Chambers

more

CoverWell ™ perfusion press-to-seal covers form water-tight, multiwell cell incubation or cytochemistry chambers when pressed to coverslips or microscope slides.…

APPLICATIONS:

Live-cell imagingMicroscopyImagingSingle molecule spectroscopy

SecureSeal™ Hybridization Chambers

more

SecureSeal™ Hybridization Chambers are thin, silicone-gasketed chambers providing optimal surface-to-volume fluid dynamics for hybridization assays on large or multiple specimens…

APPLICATIONS:

In situ hybridizationProtein and DNA MicroarraysImmunocytochemistryRapid microfluidic prototypingFluorescence Resonance Energy Transfer (FRET)

CoverWell™ Incubation Chambers

more

CoverWell™ incubation chambers are reusable, easy to apply chambers that attach without the use of adhesive.  CoverWells™ enclose a large…

APPLICATIONS:

Reverse Transfection Microarray;  DNA Microarray;  In-situ hybridization;  Immunohistochemistry; 

Silicone Isolators

more

Silicone Isolators allow researchers to isolate specimens using removable hydrophobic barriers. They may be used to isolate cells grown in…

SecureSeal™ Hybridization Chambers

more

SecureSeal™ Hybridization Chambers are thin, silicone-gasketed chambers providing optimal surface-to-volume fluid dynamics for hybridization assays on large or multiple specimens…

HybriWell™ Sealing System

more

HybriWell™ Sealing System bonds securely to a microscope slide surface in seconds to confine small reagent volumes with samples and…

Hybridization and Incubation

more

Hybridization and incubation Seals ad Chambers from Grace Bio-Labs are ideally suited for in situ-hybridization assays. The adhesive seal of…

APPLICATIONS:

In-situ hybridization MicroarraysFluorescence In situ Hybridization (FISH)FRET (Fluorescence Resonance Energy Transfer)

FastWells™ Reagent Barriers

more

FastWells™ are sticky, flexible silicone gaskets that form hydrophobic reagent barriers around specimens without messy adhesives or special slides. Gaskets may…

FlexWell™ Incubation Chambers

more

FlexWell™ incubation chamber silicone gaskets form wells on slides using clean release adhesive to isolate up to 16 specimens per…

APPLICATIONS:

Protein MicroarrayHybridizationIncubation

HybriSlip™ Hybridization Covers

more

HybriSlips™ are rigid, light-weight, thin plastic coverslips that minimize friction and facilitate uniform reagent distribution during incubation steps which require…

ProPlates®

more

ProPlates® were specifically designed to enable automated robotic liquid handling. Two main configurations are available: The ProPlate® Microtiter Plate is comprised…

APPLICATIONS:

ProteomicsProtein MicroarraysProtein expression analysis;  Antibody profiling ;  cDNA and oligonucleotide arrays; 

Silicone Isolators™ Sheet Material

more

Silicone isolator™ sheet material allows researchers to create their own removable hydrophobic barriers to isolate specimens. Where additional sealing is…

APPLICATIONS:

Protein and DNA arrays ;  Immunohistochemistry;  Fluorescence In situ Hybridization (FISH) ;  Biopolymers and hydrogel formulation ;  Cryogenic-transmission electron microscopy (Cryo-TEM) ;  Microwave crystallization ;  Ultra-small-angle X-ray scattering (USAXS) ;  Tissue ingeneering;  Live cell lithography” (LCL); 

Imaging Spacers

more

Imaging spacers are ultra-thin adhesive spacers which peel-and-stick to coverglass or microscope slides to confine specimens without compression. Layer multiple…

APPLICATIONS:

Imaging;  Microscopy;  High-temperature single-molecule kinetic analysis;  Anti‐Stokes Raman scattering microscopy; 

CoverWell™ Imaging Chambers

more

CoverWell ™ imaging chambers are designed to stabilize and support thick and free-floating specimens for confocal microscopy and imaging applications.…

APPLICATIONS:

Confocal microscopy Imaging Tissue and Cell staining ;  High Resolution Microscopy ;  Live-cell imaging ; 

CoverWell™ Perfusion Chambers

more

CoverWell ™ perfusion press-to-seal covers form water-tight, multiwell cell incubation or cytochemistry chambers when pressed to coverslips or microscope slides.…

APPLICATIONS:

Single molecule spectroscopy Live-cell imaging Microscoscopy

FastWells™ Reagent Barriers

more

FastWells™ are sticky, flexible silicone gaskets that form hydrophobic reagent barriers around specimens without messy adhesives or special slides. Gaskets may…

APPLICATIONS:

Microscopy Fluorescence In situ Hybridization (FISH) Single-molecule fluorescence analysis ;  Immunohistochemistry ; 

MultiSlip™ Coverglass Inserts

more

MutliSlip™ inserts with 8 (18mm x 18mm) or 15 (12mm x 12mm) No. 1.5 German glass coverglass per insert are…

APPLICATIONS:

High resolution microscopy Fluorescent imaging Immunohistochemistry ;  Cell Culture; 

SecureSeal™ Adhesive Sheets

more

These adhesive sheets are made using the same SecureSeal™ adhesive as is used to make HybriWell™ and SecureSeal™ Incubation Chambers.  Thin,…

APPLICATIONS:

Imaging ;  Tissue and Cell staining ;  High Resolution Microscopy; 

SecureSlip™ Silicone Supported Coverglass

more

SecureSlip™ Silicone Supported Coverglass is affixed to a thin microscopically transparent silicone base which secures it to culture vessels by…

Imaging and Microscopy

more

Imaging seals and chambers from Grace Bio-Labs offer a selection of tools for cell/tissue staining for high quality results in…

APPLICATIONS:

Tissue and Cell stainingHigh Resolution MicroscopyLive-cell imaging

CultureWell™ MultiWell Chambered Coverslips

more

CultureWell™ chambered coverglass products consist of removable and reusable, non-cytotoxic silicone gaskets secured to number 1.5 German coverglass. Chambered coverglass…

APPLICATIONS:

Cell Culture Fluorescence applications In-situ hybridization Immunostaining

CS16-CultureWell™ Removable Chambered Coverglass

more

CS16 CultureWell™ removable chambered coverglass is a 16-well chambered coverglass cell culture vessel, with 2 x 8 format with standard…

APPLICATIONS:

Cell CultureFluorescence applicationsIn-situ hybridizationImmunostaining

CultureWell™ Coverglass Inserts

more

Each CultureWell™ coverglass insert is comprised of four chambered coverglass, assembled in a disposable frame placed in a standard 86mm…

APPLICATIONS:

High resolution microscopy Fluorescent imaging Immunohistochemistry

CultureWell™ Reusable Gaskets

more

Gaskets are ideal for forming wells on glass microscope slides or in polystyrene dishes. Gaskets are non-sterile and may be…

APPLICATIONS:

Cell CultureHigh resolution microscopyFluorescent imaging Immunohistochemistry

CultureWell™ Silicone Sheet Material

more

CultureWell™ clear silicone sheet material allows researchers to create their own removable hydrophobic barriers to isolate specimens. They may be…

APPLICATIONS:

Cell CultureHigh resolution microscopy Fluorescent imagingImmunohistochemistry

MultiSlip™ Coverglass Inserts

more

MutliSlip™ inserts with 8 (18mm x 18mm) or 15 (12mm x 12mm) No. 1.5 German glass coverglass per insert are…

APPLICATIONS:

Cell CultureFluorescent imaging Immunohistochemistry

SecureSlip™ Silicone Supported Coverglass

more

SecureSlip™ Silicone Supported Coverglass is affixed to a thin microscopically transparent silicone base which secures it to culture vessels by…

APPLICATIONS:

Cell CultureImmunofluorescence assayMicroscopy

CultureWell™ ChamberSLIP 16, Non-Removable Chambered Coverglass

more

CultureWell™ NON Removable Chambered Coverglass, 16 Well, No. 1.5 German borosilicate Coverglass. Product consists of cell culture vessels, with a…

APPLICATIONS:

Cell Culture Fluorescence applicationsSmall volume incubation Immunostaining

Silicone Wound Splints

more

Wound splints are constructed of silicone and include suture sites for increased precision in affixing on or within an animal…

Silicone Isolator Sheet Material

more

Silicone isolator™ sheet material allows researchers to create their own removable hydrophobic barriers to isolate specimens. Where additional sealing is…

APPLICATIONS:

Protein and DNA arrays ;  Immunohistochemistry ;  Fluorescence In-situ Hybridization (FISH) ;  Biopolymers and hydrogel formulation;  Cryogenic-transmission electron microscopy (Cryo-TEM) ;  X-ray scattering ;  Microwave crystallization ;  Ultra-small-angle X-ray scattering (USAXS) ;  Tissue engineering Live cell lithography (LCL); 

CultureWell Silicone Sheet Material

more

CultureWell™ clear silicone sheet material allows researchers to create their own removable hydrophobic barriers to isolate specimens. They may be…

APPLICATIONS:

Lorem Ipsum ;  Lorem Ipsum;  Lorem Ipsum; 

Pathogen Genotype in Molecular Epidemiology of Waterborne Illness

IN Epoxy slides

Single nucleotide polymorphism (SNP) microarray produced using Grace Bio-Labs Epoxy Slides




The little-known parasites of the Cryptosporidium genus are responsible for the leading waterborne illness in the United States.1 The symptoms are unpleasant and an acute disease state can become life-threatening for immunocompromised individuals. In 1993, the largest recorded outbreak made 400,000 people sick in Milwaukee, WI.2 From January 2017 – March 2017, the city of Portland, OR detected Cryptosporidium in the Bull Run water source at levels requiring infrastructure overhaul.3 Cryptosporidium or “Crypto” are protected from chlorine and alcohol-based sanitizers by a thick protein-lipid-carbohydrate matrix making contaminated water difficult to treat. Construction of water filtration facilities is the only solution for Bull Run. The project is not slated for completion until 2027 and will cost an estimated $500 million US dollars.4  To better understand Cryptosporidiosis epidemiology, the Centers for Disease Control has developed a molecular tracking system, “CryptoNet”, to encourage genetic analysis of Crypto.Only molecular methods can distinguish Crypto species due to morphological similarities in clinical lab tests. Here, application of a DNA microarray using Grace Bio-Labs Epoxy Microarray Slides is employed to genotype subspecies of Crypto based on single base variations in DNA sequence, or SNPs, which occur across the genome. Known SNP variants at specific genomic loci describe a population and serve as markers to identify species, genotypes, and subtypes.6 Cyrptosporidium species and types vary in pathogenic strength, host (human vs. animal), and geographic distribution. Genetic analysis of Crypto is used to identify outbreak source (human-to-human vs. animal-to-human) and mitigate risk factors in public water supply.

Figure 1 -Genotyping assay using Grace Bio-Labs Epoxy Microarray Slides in three steps. Left panel, step 1, amino-modified oligonucleotide probes spotted on the slide surface. Middle panel, step 2, Cyrpto sample DNA was hybridized to probes. Right panel, step 3, bound sample DNA detected by Cy3 (green) or Cy5 (red) labeled

SNP Detection in Cryptosporidium Typing Array

Epoxy Microarray slides were used to distinguish C. parvum genotype I from C. parvum genotype II due to robust attachment chemistry coupled with high signal-to-noise ratio. Discrimination between five SNP variants was sufficient to unambiguously identify C. parvum genotype I, known to transmit between humans, from C. parvum genotype II, known to transmit from animals to humans.7 A SNP array is a rigorous test of microarray performance as it requires discrimination between 1 or 2 mismatches in DNA sequence. By comparison, arrays that use longer DNA based probes only need 80-85% sequence homology with sample DNA to yield a positive result.9 Two pairs of SNP probes were selected from Straub et al. 20027 to design an assay that could discriminate up to two Crypto genotype SNP variants (Figure 2). Each SNP pair contained one probe that matches genotype I DNA and one probe that matches genotype II DNA. A binary red-green output was designed as a genotyping readout. Green spots highlight DNA matching genotype I and red spots highlight DNA matching genotype II. Figure 3 shows fluorescent images of replicate arrays after hybridization and detection using a mixture of both genotype I and II Crypto DNA. Well resolved signal indicated SNP discrimination of variants from both pairs. Grace Bio-Labs ProPlate slide modules were used to optimize conditions in a high-throughput, rapid assay format. In a rapid format, DNA was hybridized for 1 hour compared with 18-hour average hybridization time in a typical assay.10 Multi-well ProPlate configurations allow replicate arrays to be assayed from the same slide surface, reducing intra-assay variation from microarray printing. Screening sample DNA concentration and hybridization conditions in multi-well ProPlates simplifies high-throughput workflows, reducing assay optimization time.

Figure 2 – Crypto genotyping assay. Replicate arrays resolve green spots of genotype I sample DNA and red spots genotype II sample DNA. Each array contains a row of control spots and four rows of SNP probes, two rows for each SNP pair. Cy3 directly labeled oligo (top row) was used as a control to show reproducible spot morphology between replicate spots. Spots were printed using non-contact piezo-electric arraying. Synthetic sample DNA was hybridized to the array followed by secondary hybridization of Cy3 and Cy5 labeled detectors. Sample DNA sequence was derived from NCBI accession AF221535.1 for genotype I modified with Iowa and GCH1 isolate SNP variants for genotype II.8

Probe Design – Number of SNP Variants Impacts Sensitivity and LOD

Performance of probes on a microarray drives assay sensitivity and the limit of detection. Factors that influence SNP probe performance include nucleotide length, position of SNP variant along the probe, number of SNP variants per probe, and modification for surface attachment.11 Two pairs of SNP probes were used to evaluate performance relative to the number of SNP variants per probe. SNP pair A contained two SNP variants per probe while SNP pair B contained one SNP variant per probe. Probe pair B had a lower limit of quantification compared to probe pair A. Both probe pairs gave a linear response to DNA concentration in support of probe design and demonstrate robust performance using Grace Bio-Labs Epoxy Microarray slides. SNP analysis on a small number of probes is shown as a genotype detection tool. A single probe to detect species or genotypes can be reliable when probes target unique regions of the genome. Rational design will allow the detection of a broad spectrum of closely related species in parallel. Epoxy microarray slides are a dependable substrate for fabrication of high-quality detection arrays. Epoxy-activated surfaces support long print runs and stable storage after printing by leveraging high reactivity and covalent immobilization of epoxy. Next-generation sequencing methods have increased the available pool of sequences to design unique diagnostic probes. While DNA sequencing remains the highest resolution genomic technique, future microarray designs will utilize newfound genetic understanding to create a user-friendly platform. Microarray technology will leverage the advantage of a high throughput, parallel, and multiplex format.

Figure 3 –SNP probes show linear response in Crypto Genotyping assay. Mean Signal-to-noise ratio (SNR) ± SD (N = 2) calculated using (signal-background) / SD background from replicate spots at each concentration of sample DNA.

enlarge table

Catalog #

Product name

405278

Epoxy Microarray Slides

246824

ProPlate® Multi-Array Slide System

244864

ProPlate® Multi-Well Chambers

470638

ProPlate® Multi-Well Chambers

106109

Vytal™ PBS

106108

Vytal™ 10 X PBS

115601

NanoParticle Fluorescent Calibration Slide

References

  1. Centers for Disease Control (CDC). (2017, November 8). Cyrptosporidiosis Surveillance, United States, 2011-2012. Morbidity and Mortality Weekly Report, Surveillance Summaries. 2015; 64(3): 1-24
  2. Gradus, S. (2014). Milwaukee, 1993: The Largest Documented Waterborne Disease Outbreak in US History. Retrieved from https://waterandhealth.org/safe-drinking-water/drinking-water/milwaukee-1993-largest-documented-waterborne-disease-outbreak-history/
  3. Portland Water Bureau, City of Portland, OR. (2018). Information on Cryptosporidium. Retrieved from https://www.portlandoregon.gov/water/75112
  4. Portland Water Bureau, City of Portland, OR (2018). Bull Run LT2 Interim Measures Watershed Inspection and Monitoring Plan. Retrieved from https://www.portlandoregon.gov/water/article/669108
  5. Centers for Disease Control and Prevention (CDC). (2015, January 28). CryptoNet: Molecular-based Tracking to Better Understand U.S. Cryptosporidiosis Transmission. Retrieved from https://www.cdc.gov/parasites/crypto/cryptonet.html
  6. Kitts, A. et al. (2014). The Database of Short Genetic Variation (dbSNP). Retrieved from https://www.ncbi.nlm.nih.gov/books/NBK174586/
  7. Straub TM, Daly DS, Wunshel S, Rochelle PA, DeLeon R, Chandler DP. Genotyping Cryptosporidium parvumwith an hsp70 Single-Nucleotide Polymorphism Microarray. Applied and Environmental Microbiology. 2002;68(4):1817-1826. doi:10.1128/AEM.68.4.1817-1826.2002.
  8. Peng, M.M. et al. Genetic polymorphism Among Cryptosporidium parvum Isolates: Evidence of Two Distinct Human Transmission Cycles. Emerging Infectious Diseases. Centers for Disease Control. October – December 1997; 3(4): 567-573.
  9. Kostić, T. and Sessitsch, A. Microbial Diagnostic Microarrays for the Detection and Typing of Food-and Water-Borne (Bacterial) Pathogens. Microarrays. 2012; 1:3-24. doi:10.3390/microarrays1010003
  10. Chagovetz, A. and Blair, S. Real-time DNA microarrays: reality check. Biochem Soc Trans. 2009 April; 37(Pt 2): 471-475. doi: 10.1032/BST0370471
  11. Dufva, M. Fabrication of high quality microarrays. Biomolecular Engineering. 22(5-6):173-184. doi: 10.1016/j.bioeng.2005.09.003
  12. Miller, M.B. and Tang Y. (2009). Basic Concepts of Microarrays and Potential Applications in Clinical Microbiology. Clinical Microbiology Reviews. 2009; 22(4): 611-633.
  13. Li, B et al. Advancements in Microarray Utility for Detection and tracking of Foodborne Microbes in the Genomic Era. Advanced Techniques in Biology and Medicine. 2017; 5(3). Doi: 10.4172/2379-1764.1000239.