Evidence map›Paper›PMID 32283940›Full record

ArticleBioTechniques2020

Optimization of western blotting for the detection of proteins of different molecular weight.

Ghanshyam D Heda, Lisa Shrestha, Sagarina Thapa, Shreya Ghimire, Diptika Raut

Open access · goldAbstract read
In one paragraph

Article in BioTechniques, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
0.6field-weighted citation impact, top 37% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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.

2 · The registry

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.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 11 citations in OpenAlex.

  1. Article
  2. Article
  3. Review
  4. Comprehensive Optimization of Western Blotting.Gels (Basel, Switzerland) · 2023
    Article
  5. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors at 3 institutions in 1 country.

Ghanshyam D HedaDepartment of Sciences & Mathematics, Mississippi University for Women, Columbus, MS 39701, USA.
Lisa ShresthaDepartment of Sciences & Mathematics, Mississippi University for Women, Columbus, MS 39701, USA.
Sagarina ThapaDepartment of Sciences & Mathematics, Mississippi University for Women, Columbus, MS 39701, USA.
Shreya GhimireDepartment of Sciences & Mathematics, Mississippi University for Women, Columbus, MS 39701, USA.
Diptika RautDepartment of Sciences & Mathematics, Mississippi University for Women, Columbus, MS 39701, USA.
Mississippi University for Women · USUniversity of Iowa · USUniversity of Maryland, Baltimore · US

Funding

Training and Mentoring Core P20GM103476 · NIGMS · UNIVERSITY OF SOUTHERN MISSISSIPPI · PI MICHAEL R GARRETT · 2012 to 2026
$60.2M
NIGMS NIH HHS P20 GM103476
6 · The paper itself

Abstract

Protein samples electroblotted onto nitrocellulose membranes and quenched with a mixture of blocking agents produced a strong signal for cystic fibrosis transmembrane-conductance regulator (CFTR), a high-molecular-weight protein, in western blotting. Optimized conditions for CFTR were then extended to medium- and low-molecular-weight proteins (LAMP1 and Rab11a, respectively) to determine the effects of methanol concentration (0-20%) in Towbin's transfer buffer (TTB). Methanol in TTB appears to have little to no effect on CFTR signal. However, for medium-sized (LAMP1) and small (Rab11a) proteins, a lower concentration of methanol (10%) was sufficient to produce a maximal signal. Therefore, methanol, a toxic solvent, can be removed from or reduced in TTB without compromising signal strength. Here, we show modifications that may be useful in detecting and/or improving the signal of low-abundance proteins.

Indexed as

Blotting, WesternCell LineCollodionCystic Fibrosis Transmembrane Conductance RegulatorHumansMolecular WeightSignal TransductionTransfectionCFTR protein, humanCollodionCystic Fibrosis Transmembrane Conductance Regulatorblocking agentsCFTRLAMP1methanol concentrationoptimizationRab11aTowbin’s transfer bufferwestern blotting

Identifiers

PMID32283940
PMCPMC7333534
OpenAlexW3015809021

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

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.