ArticleJournal of visualized experiments : JoVE2016
Protein Complex Affinity Capture from Cryomilled Mammalian Cells.
Article in Journal of visualized experiments : JoVE, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
26 citing papers in PubMed, 36 citations in OpenAlex.
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- O-Mannose Glycosylations Influence E-Cadherin Functional Interactions.Molecular & cellular proteomics : MCP · 2026Article
- Proteomics reveals extensive phosphoregulation of outer kinetochore protein KNL1.bioRxiv : the preprint server for biology · 2026Article
- Proteomics reveals extensive phosphoregulation of outer kinetochore protein KNL1.microPublication biology · 2026Article
- Repeats mimic pathogen-associated patterns across a vast evolutionary landscape.Cell genomics · 2025Article
- Article
- Microbial iCLIP2: enhanced mapping of RNA-protein interaction by promoting protein and RNA stability.RNA (New York, N.Y.) · 2025Article
- Cancer cells restrict immunogenicity of retrotransposon expression via distinct mechanisms.Immunity · 2024Article
- Multiparameter screen optimizes immunoprecipitation.BioTechniques · 2024Article
- CCDC88B interacts with RASAL3 and ARHGEF2 and regulates dendritic cell function in neuroinflammation and colitis.Communications biology · 2024Article
- 7SK methylation by METTL3 promotes transcriptional activity.Science advances · 2023Article
- Structure and dynamics of the essential endogenous mycobacterial polyketide synthase Pks13.Nature structural & molecular biology · 2023Article
- Proteomic elucidation of the targets and primary functions of the picornavirus 2A protease.The Journal of biological chemistry · 2022Article
- Affinity Isolation of Endogenous Saccharomyces Cerevisiae Nuclear Pore Complexes.Methods in molecular biology (Clifton, N.J.) · 2022Article
- MeasuringSTAR protocols · 2021Article
- Affinity proteomic dissection of the human nuclear cap-binding complex interactome.Nucleic acids research · 2020Article
- Diversification of CORVET tethers facilitates transport complexity inJournal of cell science · 2020Article
- LINE-1 ORF2p expression is nearly imperceptible in human cancers.Mobile DNA · 2020Article
- Identification of RNase-sensitive LINE-1 Ribonucleoprotein Interactions by Differential Affinity Immobilization.Bio-protocol · 2019Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Affinity capture is an effective technique for isolating endogenous protein complexes for further study. When used in conjunction with an antibody, this technique is also frequently referred to as immunoprecipitation. Affinity capture can be applied in a bench-scale and in a high-throughput context. When coupled with protein mass spectrometry, affinity capture has proven to be a workhorse of interactome analysis. Although there are potentially many ways to execute the numerous steps involved, the following protocols implement our favored methods. Two features are distinctive: the use of cryomilled cell powder to produce cell extracts, and antibody-coupled paramagnetic beads as the affinity medium. In many cases, we have obtained superior results to those obtained with more conventional affinity capture practices. Cryomilling avoids numerous problems associated with other forms of cell breakage. It provides efficient breakage of the material, while avoiding denaturation issues associated with heating or foaming. It retains the native protein concentration up to the point of extraction, mitigating macromolecular dissociation. It reduces the time extracted proteins spend in solution, limiting deleterious enzymatic activities, and it may reduce the non-specific adsorption of proteins by the affinity medium. Micron-scale magnetic affinity media have become more commonplace over the last several years, increasingly replacing the traditional agarose- and Sepharose-based media. Primary benefits of magnetic media include typically lower non-specific protein adsorption; no size exclusion limit because protein complex binding occurs on the bead surface rather than within pores; and ease of manipulation and handling using magnets.
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Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.