Protoarray human protein microarrays v5

Manufactured by Thermo Fisher Scientific

Sourced in United States

The ProtoArray® Human Protein Microarrays v5.0 is a comprehensive collection of human proteins printed on a glass slide. It allows for the analysis of protein-protein, protein-small molecule, and protein-antibody interactions.

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Lab products found in correlation

Quadriperm dishes, by Greiner (1 mentions) Axon 4200al, by Molecular Devices (1 mentions) Alexa fluor 647 conjugated goat anti human igg, by Thermo Fisher Scientific (1 mentions) Genepix pro software, by Molecular Devices (1 mentions) Axon genepix 4000b, by Molecular Devices (1 mentions)

Close spelling variants of this product

ProtoArray® Human Protein Microarray v5.0 ProtoArray Human Protein Microarrays Human ProtoArray v5.0 Protoarray® v5.0 ProtoArray

4 protocols using protoarray human protein microarrays v5

Protein-Protein Interaction Profiling

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EXAMPLE 8

Invitrogen's Protein-Protein Interaction Profiling Service was performed on ProtoArray® Human Protein Microarrays v5.0 (protein microarray) and the interaction with more than 9,000 human proteins was investigated. One Alexa Fluor® (fluorescent dye) 647-conjugated synthetic peptide, provided by the CARI Reproductive Institute, designated as PIF-Alexa 647, was used to probe the microarrays at two concentrations (250 nM and 2.5 μM) in duplicate. The results from these assays were compared to a negative control assay in which the probe protein was excluded. All interactions of the probe protein with the immobilized proteins on the microarrays were evaluated by specified statistical threshold criteria within the array and compared to interactions observed on the negative control array. Visual confirmation of the positive hits was also performed by directly inspecting the array images to verify the authenticity and quality of the signals. From these analyses, a total of eleven (11) human proteins were identified as candidate interactors for PIF-Alexa 647 (fluorescent dye).

US10953071B2. Compositions and methods for treatment of intracellular damage and bacterial infection (2021-03-23). BioIncept, LLC [US]. Inventors: Eytan R. Barnea [US].

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Protein Microarray Profiling of Sera

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ProtoArray Human Protein Microarrays v5.0 (Invitrogen, Carlsbad, CA, USA) were purchased and used according to the manufacturer’s instructions.
After blocking for 1 h at 4 °C and washing, arrays were incubated in quadriPERM dishes (Greiner Bio-One, Frickenhausen, Germany) placed on a horizontal shaker (50 rpm) for 90 min at 4 °C with individual sera diluted at 1:500 in 5 mL washing buffer (0.1% Tween 20 [v/v], 1% BSA [w/v] in PBS). After washing, binding of IgG was detected by incubation with Alexa Fluor 647–conjugated goat anti–human IgG (Invitrogen, Waltham, MA, USA) diluted 1:2000 in assay buffer for 90 min at 4 °C. The arrays were washed again and dried by centrifugation. Arrays were scanned at a 10-μm resolution on a microarray scanner (Axon 4200AL with GenePix Pro Software; Molecular Devices, Sunnyvale, CA, USA), and fluorescence was detected according to the manufacturer’s instructions. Images were saved as 16-bit TIFF files, and analysis was performed using GenePix. The median net intensity in relative fluorescence units is reported for each spot.

Hamano Y., Kida H., Ihara S., Murakami A., Yanagawa M., Ueda K., Honda O., Tripathi L.P., Arai T., Hirose M., Hamasaki T., Yano Y., Kimura T., Kato Y., Takamatsu H., Otsuka T., Minami T., Hirata H., Inoue K., Nagatomo I., Takeda Y., Mori M., Nishikawa H., Mizuguchi K., Kijima T., Kitaichi M., Tomiyama N., Inoue Y, & Kumanogoh A. (2017). Classification of idiopathic interstitial pneumonias using anti–myxovirus resistance-protein 1 autoantibody. Scientific Reports, 7, 43201.

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Profiling Protein Interactions Using PIF-Alexa Microarray

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ProtoArray Human Protein Microarrays v5.0 (Invitrogen, Carlsbad, CA) having 9000 possible targets were initially blocked and then exposed to PIF-Alexa 647 (250 nM and 2.5 µM). Unbound probe was washed off and arrays read using a fluorescence microarray scanner. As a negative control, an array was incubated with wash buffer while calmodulin kinase was used as positive control detected by AlexaFluor 647-conjugated anti-v.5 antibody. Three stringent conditions determined AlexaFluor-PIF specificity of binding; signal intensity >200 relative fluorescent units, >6 time higher than the negative control, and Z-factor >5.

Barnea E.R., Lubman D.M., Liu Y.H., Absalon-Medina V., Hayrabedyan S., Todorova K., Gilbert R.O., Guingab J, & Barder T.J. (2014). Insight into PreImplantation Factor (PIF*) Mechanism for Embryo Protection and Development: Target Oxidative Stress and Protein Misfolding (PDI and HSP) through Essential RIPK Binding Site. PLoS ONE, 9(7), e100263.

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Serum Protein Profiling Using ProtoArray

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Serum samples from pre- (week 0) and the latest post-treatment time point were profiled on ProtoArray® Human Protein Microarrays v5.0 (Invitrogen, Carlsbad, CA) containing approximately 9000 human proteins. One ProtoArray slide was used for each time point and slides were processed according to the manufacturer’s instructions and scanned using an Axon GenePix 4000B fluorescent microarray scanner (Molecular Devices, Sunnyvale, CA).

Sanborn R.E., Ross H.J., Aung S., Acheson A., Moudgil T., Puri S., Hilton T., Fisher B., Coffey T., Paustian C., Neuberger M., Walker E., Hu H.M., Urba W.J, & Fox B.A. (2017). A pilot study of an autologous tumor-derived autophagosome vaccine with docetaxel in patients with stage IV non-small cell lung cancer. Journal for Immunotherapy of Cancer, 5, 103.

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