ArticleBioTechniques2022
MILKSHAKE: novel validation method for antibodies to post-translationally modified targets by surrogate Western blot.
Article in BioTechniques, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 8 citations in OpenAlex.
- Sprinkling in extra validation for high-value PTMs and therapeutic Abs with MILKSHAKE Western blots and Sundae ELISAs.New biotechnology · 2025Article
- Site-directed neutralizing antibodies targeting structural sites on SARS-CoV-2 spike protein.New biotechnology · 2024Article
- Proteomics-The State of the Field: The Definition and Analysis of Proteomes Should Be Based in Reality, Not Convenience.Proteomes · 2024Article
- Engineered FHA domains can bind to a variety of Phosphothreonine-containing peptides.Protein engineering, design & selection : PEDS · 2024Article
- MILKSHAKE Western blot and Sundae ELISA: We all scream for better antibody validation.Journal of immunological methods · 2023Article
- Validation and the Determination of Antibody Bioactivity Using MILKSHAKE and Sundae Protocols.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Epivolve: A Protocol for Site-Directed Antibodies.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Benchmarking antibody discovery fidelity and reproducibility with an assay-locked residue fidelity index.BioTechniquesArticle
Corrections and comments
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Authors and funding
11 authors at 2 institutions in 1 country.
Funding
Abstract
Antibody (Ab) validation is the procedure in which an Ab is thoroughly assayed for sensitivity and specificity in a given application. Validation of Abs against post-translationally modified (PTM) targets is particularly challenging because it requires specifically prepared antigen. Here we describe a novel validation method using surrogate proteins in a Western blot. The surrogate protein, which we termed 'MILKSHAKE,' is a modified maltose binding protein enzymatically conjugated to a peptide from the chosen target that is either modified or nonmodified at the residue of interest. The certainty of the residue's modification status can be used to confirm Ab specificity. This method also allows for Ab validation even in the absence or limited availability of treated cell lysates.
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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.