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agarose streptavidin  (Vector Laboratories)
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Vector Laboratories agarose streptavidin
Mass spectrometry identifies nuclear paraspeckles as interacting partners of unphosphorylated miR-34 probes. ( A ) Workflow of sample isolation for mass spectrometry. <t>Streptavidin</t> sepharose beads were incubated with biotinylated miRNA mimics and mixed with sonicated whole-cell lysates. Samples were eluted by RNAse A treatment. The figure was created with Biorender. ( B ) Curation of mass spectrometry hits. Candidate proteins were identified by significance, uniqueness to the 5′OH miR-34a probe, and presence of known RNA-binding activity. ( C ) Characterization of isolated proteins. The total number of proteins isolated binding to the indicated probe is shown in parentheses, with the number of proteins unique to that sample or overlapping with other samples shown in the Venn diagram. ( D ) Gene ontology (GO) curation for cellular compartments performed on proteins identified in the 5′OH miR-34a eluates before and after curation (which was the removal of hits with a score below 20, p value > 0.05, non-unique proteins across three replicates, and non-RNA-binding proteins).
Agarose Streptavidin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Mass spectrometry identifies nuclear paraspeckles as interacting partners of unphosphorylated miR-34 probes. ( A ) Workflow of sample isolation for mass spectrometry. Streptavidin sepharose beads were incubated with biotinylated miRNA mimics and mixed with sonicated whole-cell lysates. Samples were eluted by RNAse A treatment. The figure was created with Biorender. ( B ) Curation of mass spectrometry hits. Candidate proteins were identified by significance, uniqueness to the 5′OH miR-34a probe, and presence of known RNA-binding activity. ( C ) Characterization of isolated proteins. The total number of proteins isolated binding to the indicated probe is shown in parentheses, with the number of proteins unique to that sample or overlapping with other samples shown in the Venn diagram. ( D ) Gene ontology (GO) curation for cellular compartments performed on proteins identified in the 5′OH miR-34a eluates before and after curation (which was the removal of hits with a score below 20, p value > 0.05, non-unique proteins across three replicates, and non-RNA-binding proteins).

Journal: Non-Coding RNA

Article Title: Paraspeckles Are Associated with the Activation and Nuclear Localization of Unphosphorylated miR-34a

doi: 10.3390/ncrna12020012

Figure Lengend Snippet: Mass spectrometry identifies nuclear paraspeckles as interacting partners of unphosphorylated miR-34 probes. ( A ) Workflow of sample isolation for mass spectrometry. Streptavidin sepharose beads were incubated with biotinylated miRNA mimics and mixed with sonicated whole-cell lysates. Samples were eluted by RNAse A treatment. The figure was created with Biorender. ( B ) Curation of mass spectrometry hits. Candidate proteins were identified by significance, uniqueness to the 5′OH miR-34a probe, and presence of known RNA-binding activity. ( C ) Characterization of isolated proteins. The total number of proteins isolated binding to the indicated probe is shown in parentheses, with the number of proteins unique to that sample or overlapping with other samples shown in the Venn diagram. ( D ) Gene ontology (GO) curation for cellular compartments performed on proteins identified in the 5′OH miR-34a eluates before and after curation (which was the removal of hits with a score below 20, p value > 0.05, non-unique proteins across three replicates, and non-RNA-binding proteins).

Article Snippet: Lysates were cleared by incubating overnight with 100 μL agarose streptavidin (Vector, Malvern, PA, USA SA-5010).

Techniques: Mass Spectrometry, Isolation, Incubation, Sonication, RNA Binding Assay, Activity Assay, Binding Assay