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In the world of science, citations are currency. They signal impact, influence, and utility. Ask most people to name the most important biology paper ever, and you might hear Watson & Crick's 1953 DNA double helix (a landmark, but with "only" tens of thousands of citations). Yet the actual record-holder? A 1951 method for measuring protein concentration.
As of 2025–2026 data from Web of Science and Nature's April 2025 update, biology's citation kings are overwhelmingly methodological papers — simple, reliable protocols that every lab uses daily. These aren't flashy discoveries; they're workhorse tools. Researchers cite them routinely: "Protein was quantified using Lowry et al. (1951)."
Why? Biology is experimental. Every gel, blot, extraction, or assay needs a reference. Breakthrough ideas often become textbook facts and get cited less directly. Methods papers rack up citations for decades.
The Undisputed Champion: Lowry et al. (1951)
Title: Protein measurement with the Folin phenol reagent Authors: Oliver H. Lowry, Nira J. Rosebrough, A. Lewis Farr, Rose J. Randall Journal: Journal of Biological Chemistry 193: 265–275 [Read PDF] Citations: >350,000–355,000+ (still #1 across all science per Nature 2025)
This humble assay uses copper and Folin-Ciocalteu reagent to detect proteins via color change. It's sensitive, cheap, and reproducible. Even with alternatives like Bradford (1976), labs worldwide still rely on it or cite it as the classic standard.
Oliver Lowry himself was surprised by its runaway success. In later reflections, he noted it wasn't his "greatest paper," yet he enjoyed the fame. Today, it gains thousands of citations yearly.
The Methods Hall of Fame: Top Cited in Biology/Biochemistry
Here's a snapshot of the perennial top contenders (based on WoS/Nature 2025 data, GenScript compilations, and recent cross-checks). Citation counts are approximate and continue climbing.
| Rank | Paper (Year) | Authors | Journal | Approx. Citations (WoS) | What It Does |
|---|---|---|---|---|---|
| 1 | Protein measurement with the Folin phenol reagent (1951) | Lowry et al. | J. Biol. Chem. | >350,000 | Classic protein quantification assay |
| 2 | Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) | Laemmli | Nature | >300,000 | SDS-PAGE: Gold-standard protein gel electrophoresis |
| 3 | A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) | Bradford | Anal. Biochem. | >200,000 | Bradford (Coomassie) assay — faster Lowry alternative |
| 4 | Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction (1987) | Chomczynski & Sacchi | Anal. Biochem. | >100,000–150,000 | RNA extraction (basis for TRIzol) |
| 5 | Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications (1979) | Towbin et al. | PNAS | >100,000 | Western blotting technique |
| 6 | DNA sequencing with chain-terminating inhibitors (1977) | Sanger et al. | PNAS | High tens of thousands | Sanger dideoxy sequencing method |
| 7 | High-resolution two-dimensional electrophoresis of proteins (1975) | O'Farrell | J. Biol. Chem. | High tens of thousands | 2D gel electrophoresis for protein separation |
Influential Reviews That Shape Thinking
Methods lead, but conceptual reviews can accumulate massive citations too:
- The hallmarks of cancer (Hanahan & Weinberg, 2000 & 2011 updates in Cell) — >100,000+ combined. Frames modern cancer research.
- Hallmarks of aging (López-OtÃn et al., various updates, e.g., 2023 in Cell) — Rapidly rising, especially the 2023 version with 12 hallmarks.
These synthesize ideas and become go-to references.
The Modern Shift: Faster Accumulators
All-time lists favor old methods, but recent papers (post-2000) climb quickly:
- CRISPR-Cas9 papers (e.g., Jinek et al. 2012; Cong et al. 2013) — Tens of thousands already.
- AlphaFold papers (Jumper et al. 2021; Abramson et al. 2024 on AlphaFold 3) — Explosive growth; AlphaFold 3 already has thousands in short time.
- Single-cell omics, deep learning in biology, and aging reviews.
In 2025 Google Scholar metrics, tools like AlphaFold and new AI-biology hybrids are rising stars.
Why Biology Citations Look Like This (Unlike Physics or Math)
In physics, theoretical breakthroughs (e.g., quantum mechanics papers) often top lists. Biology rewards utility. A great idea might inspire 1,000 papers; a great protocol gets cited in 100,000 experiments.
This reflects biology's hands-on nature — and why technicians, students, and PIs all cite the same classics.
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