Category: Featured Research

Optimized Blocking Of Porous Nitrocellulose Films For Sensitive Protein Microarrays

Michael A. Shultz, Aki Ohdera, Jason MacManiman, Charles M. McGrath. Grace Bio-Labs, Inc.   ABSTRACT Super G Blocking Buffer was developed to produce low backgrounds in assays using nitrocellulose film-slides.  Using this blocking reagent in combination with ONCYTE® SuperNOVA Film-slides, we achieved an order of magnitude greater sensitivity in a cytokine microarray assay than with … Continued

Inclusion of Super G in All Steps of Protein Arrays Improves S/N

Blocking reagents are used to reduce non-specific protein binding in immunoassays, significantly improving results in terms of Signal-to-Noise ratio. Typically the blocking reagent is added prior to addition of the primary detection antibody.In this study we asked the question as to whether addition of the blocking reagent to other steps in a protein array assay … Continued

Optimizing Reverse Phase Protein Array Assays

Components of Reverse-Phase Protein Array (RPPA) methods and their impact on the overall performance of this assay were presented at the 2nd annual RPPA Conference in Edinburgh, Scotland on November 12.  Entire presentation can be viewed here.  View poster here. View here for information regarding our RPPA Assay System Selecting Surfaces, Reagents, and Detection Methods … Continued

Preserving Protein Immuno-Reactivity

Microarrays deposited onto solid supports are robust platforms for performing antibody capture and antigen detection assays.  Among various substrates, thin film porous nitrocellulose cast on glass surfaces has excelled in the field of proteomics since its invention by scientists at Grace Bio-Labs (McGrath, CM, Grudzien, J and A. Levine. Cell Vision 2:499-509, 1995). The irregular 3-dimensional … Continued

Conserve Reagents with Miniature Western Blot Reaction Chambers

SecureSeal Hybridization Chambers were originally developed to perform hybridizations on specimens affixed to glass slides or to perform microarray incubations with limited volumes (SecureSeal References). The peel- and- stick adhesive chambers can be created in various sizes to accommodate different samples, including partial membrane blots from protein mini-gels (shown here).  Recently we have investigated the … Continued

From Western Blot to Protein Array with Super G Blocking Buffer

Results obtained from Western blots can be highly variable and dependent not only on the specific antigen/antibody pairs used, but also on other reagents in the protocol such as blocking buffer. Not surprisingly, no one “universal” blocking reagent works well with all proteins. Thus choice of blocking reagent should be assessed for optimal results and having … Continued

View Your Proteins in 3-D!

Protein microarray technology has progressed significantly and, although new microarray surface chemistries continue to be developed and marketed, porous nitrocellulose has remained the predominant immobilization surface for protein microarray applications due to its many functional advantages.  The 3-dimensional structure of nitrocellulose film slides offers a considerably higher surface area for protein binding compared to conventional … Continued

ProPlate® Chambers Minimize Microarray Variation

Microarray results are susceptible to variation introduced from multiple sources during the course of assay methodologies employed with this technology. A commonly overlooked parameter which may significantly impact results is the choice of chamber/vessel for performing microarray incubations and washes. Chambers which allow for active mixing can significantly improve microarray data. Grace Bio-Labs offers ProPlate® incubation chambers which allow for continuous mixing during microarray assays … Continued

Protein Staining Nitrocellulose using SYPRO® Ruby

Reverse phase protein arrays (RPPA) offer great promise for the identification and analysis of critical biomarkers for disease states in patient samples and for high-throughput drug screening using cultured cell systems.  Porous nitrocellulose (NC) film slides continue to be the preferred substrate for RPPA due to their high protein binding capacity and many functional advantages. … Continued

SuperNOVA: Superior Porous Nitrocellulose Surface for Fluorescence Detection

Fluorescence detection is a preferred method for many quantitative protein microarray assays in research and diagnostic applications due to its excellent sensitivity and potential for multiplexing.  Use of multiplexed fluorescence-based assays on porous nitrocellulose though has been hampered by high backgrounds at shorter excitation wavelengths (such as the 532 nm fluorescence channel). We have developed … Continued

Super G: Super Blocking and Signal-to-Noise for ONCYTE Film Slides

Fluorescence detection is a preferred method for many quantitative protein microarray assays in research and diagnostic applications due to its excellent sensitivity and potential for multiplexing. Use of multiplexed fluorescence-based assays on porous nitrocellulose though has been hampered by high backgrounds at shorter excitation wavelengths (such as the 532 nm fluorescence channel).  To maximize the … Continued

Print Macroarrays with ProPlate

ProPlate® Chambers for Macroarray Printing Many laboratories desire the benefits of utilizing microarray technology in their field of interest but do not have access to microarray printing equipment or cannot justify the expense of dedicated print runs for lower throughput experiments.  In many cases where highly multiplexed assays are not a necessity or during early-phase … Continued