Protein Blotting of Basic Proteins Resolved on Acid-Urea-Triton-Polyacrylamide Gels
Bibliographic record
Abstract
The electrophoretic resolution of histones on acetic acid-urea-Triton (AUT) polyacrylamide gels is the method of choice to separate basic proteins, such as histone variants, modified histone species, and high-mobility group proteins 14 and 17 ( 1 - 6 and see Chapters 16 and 17 ). Basic proteins are resolved in this system on the basis of their size, charge, and hydrophobicity. In previous studies, we analyzed the abundance of ubiquitinated histones by resolving the histones on two-dimensional (AUT into SDS) polyacrylamide gels, followed by their transfer to nitrocellulose membranes, and immunochemical staining of nitrocellulose membranes with an antiubiquitin antibody ( 7 - 9 ). However, transfer of the basic proteins directly from the AUT polyacrylamide gel circumvents the need to run the second-dimension SDS gel and accomplishes the analysis of several histone samples. We have described a method that efficiently transfers basic proteins from AUT polyacrylamide gels to nitrocellulose membranes ( 10 ). This method has been used in the immunochemical detection of modified histone, isoforms, and histone H1 subtypes ( 6 , 11 - 13 ). These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.008 | 0.009 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".