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- Efficiently produce up to 12 differentiated subproteomes with uncompromised functional and structural integrity
- Generate characteristic molecular profiles for comparison and discovery
- Enrich functional biomarkers for sequence and structural annotation
- Kit includes 6 mixed mode surface chemistries per prep
Functional proteomics relies in part, on the functional or structural features of intact, non-denatured proteins. While the terminology can often overlap, chemical and affinity-based proteomic profiles can be considered a subset of functional proteomics. Thus, functional proteomic annotation may complement conventional sequence annotation while supporting the study of mechanism of action and drug promiscuity. Furthermore, the subtleties of protein attributes, when the same or similar underlying sequence can have multiple conformations and functions, and when different sequences sometime perform the same or similar functions, are now open to investigation. A
new functional proteomics separations toolset based on the NuGel™ passivated porous silica platform, can be used in unrestricted workflow strategies, so as to sift through these biological complexities.
- Separations readily compatible with virtually all proteomic interrogations
- Microtube kit, simple bind/wash/elute protocols
- No specialized instruments, or HPLC required
- Disposable, no column regeneration
- Tryptic digestion or enzyme assay can be ‘on-bead’
- Universal, species and tissue type agnostic
Click Here To View NuGel™ NRicher™ 6 Product Sheet
Mutations, post-translational modification, and non-covalent binding factors all play a role in fine tuning polypeptide sequence to final function. Because of this phenomena, populations of proteins annotated to the same sequence nevertheless can display multiple functions within tissues and disease states. Likewise, the same or similar function may be presented by multiple sequences, with subcellular control mechanisms regulating functional diversity. As a consequence, strictly abundance based biomarkers may lack the necessary dynamic range and greater specificity provided by functional based biomarkers, to define the phenotype. Thus, Functional Proteomic techniques such as described here, support a top-down proteomic strategy starting with functional annotation of the structurally intact protein, and ending with sequence and structural annotation.
Two of BSG products, NRicher™ 6 and HemogloBind™, were able to contribute to this rigorous examination of protein complexes. When our products were used as a pretreatment step in the overall workflow, about twice the number of observations and annotations became possible. This further validates that the sub-proteome bias characteristics of NRicher™ 6 can simplify complex proteomes into less complex sub-proteomes with efficiencies suitable for deep functional proteome characterization. Furthermore, this study demonstrated the importance of a key feature implicit to all of our products; that is the maintenance of functional and structural integrity after separations. Without that particular feature, these additional observations would not have been possible.
The surface chemisties reported here as SeraFILE in this article form the basis of the product NRicher™ 6.
Functional proteomic profiling can help identify targets for disease diagnosis and therapy. Available methods are limited by the inability to profile many functional properties measured by enzymes kinetics. The functional proteomic profiling approach proposed here seeks to overcome such limitations. It begins with surface-based proteome separations of tissue/cell extracts, using SeraFILE, a proprietary protein separations platform. Enzyme kinetic properties of resulting subproteomes are then characterized, and the data integrated into proteomic profiles. As a model, SeraFILE-derived subproteomes of cyclic nucleotide-hydrolyzing phosphodiesterases (PDEs) from bovine brain homogenate (BBH) and rat brain homogenate (RBH) were characterized for cAMP hydrolysis activity in the presence (challenge condition) and absence of cGMP. Functional profiles of RBH and BBH were compiled from the enzyme activity response to the challenge condition in each of the respective subproteomes. Intersample analysis showed that comparable profiles differed in only a few data points, and that distinctive subproteomes can be generated from comparable tissue samples from different animals. These results demonstrate that the proposed methods provide a means to simplify intersample differences, and to localize proteins attributable to sample-specific kinetic responses. It can be potentially applied for disease and non-disease sample comparison in biomarker discovery and drug discovery profiling.
Kit includes:
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Kit Contains:
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NuGel™ NRicher™ 6
5 Preps
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NuGel™ NRicher™ 6
25 Preps
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PRO- (A,B,C,L,N,R) reagent powders
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75 mg each reagent
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375 mg each reagent
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PRO-BB Binding Buffer, pH 6.0
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15 ml
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75 ml
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PRO-EB Elution Buffer, pH 9.0
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1.5 ml
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7.5 ml
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Spin-X microfuge filters
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30
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150
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MSDS:
MSDS - NuGel™ NRicher™ 6, Catalog Order # SR610-5, SR610-25
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References
"NRicher™: A Singular Low Abundance Proteome Enrichment Product Line With Seamless Integration of On-Bead Digestion " Biotech Support Group to exhibit at the 71st ASMS Conference on Mass Spectrometry and Allied Topics, June 4 - 8, 2023 in Houston, Texas
C Wan, B Borgeson, S Phanse, F Tu, K Drew, G Clark, et al. Panorama of ancient metazoan macromolecular complexes. Nature Volume:525, Pages:339–344 Date published:(17 September 2015). doi:10.1038/nature14877 The Functional & Chemical Proteomics Handbook 03/2016
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