Evidence map›Paper›PMID 31672165›Full record

ArticleBMC research notes2019

Contaminating reactivity of a monoclonal CCAAT/Enhancer Binding Protein β antibody in differentiating myoblasts.

Hamood AlSudais, Nadine Wiper-Bergeron

Open access · goldAbstract read
In one paragraph

Article in BMC research notes, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
0.1field-weighted citation impact, top 51% 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

2 citing papers in PubMed, 2 citations in OpenAlex.

  1. Article
  2. 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

2 authors at 1 institution in 1 country.

Hamood AlSudaisGraduate Program in Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Rm 3106, K1H 8M5, Ottawa, ON, Canada.
Nadine Wiper-BergeronDepartment of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Rm 2539, K1H 8M5, Ottawa, ON, Canada. Nadine.WiperBergeron@uottawa.ca.ORCID http://orcid.org/0000-0001-8448-3246
University of Ottawa · CA

Funding

CIHR 148613
6 · The paper itself

Abstract

objectiveCCAAT/Enhancer Binding proteins (C/EBPs) are transcription factors involved in the regulation of a variety of cellular processes. We used the Abcam Recombinant Anti-C/EBP beta antibody (E299) to detect C/EBPβ expression during myogenesis. Though the antibody is monoclonal, and the immunogen used is highly specific to C/EBPβ, we identified an intense band at 23 kDa on western blot that did not correspond to any of the known isoforms of C/EBPβ, or family members predicted to cross-react. Absent in myoblast cells overexpressing C/EBPβ, the band was present when C/EBPβ was knocked down, confirming specificity for a protein other than C/EBPβ. The objective of this work was to identify the contaminating reactivity.

resultsWe performed immunoprecipitation followed by mass spectrometry to identified myosin light chain 4 (MYL4) as the unknown band, suggesting that the Abcam monoclonal antibody directed against C/EBPβ is not pure, but contains a contaminating antibody against MYL4. Caution should be used when working in cells lines that express MYL4 to not confound the detection of MYL4 with that of C/EBPβ isoforms.

Indexed as

AnimalsAntibodies, MonoclonalAntibody SpecificityCCAAT-Enhancer-Binding Protein-betaCell DifferentiationCell LineCross ReactionsGene Expression ProfilingHEK293 CellsHumansMiceMuscle DevelopmentMyoblastsMyosin Light ChainsProtein IsoformsTandem Mass SpectrometryAntibodies, MonoclonalCCAAT-Enhancer-Binding Protein-betaCEBPB protein, humanMYL4 protein, humanMyosin Light ChainsProtein IsoformsAntibody cross-reactivityC/EBPβMYL4MyoblastMyogenesis

Identifiers

PMID31672165
PMCPMC6824081
OpenAlexW2982859365

What OpenQuestion holds

Textmetadata
LicenceCC BY
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.