Evidence map›Paper›PMID 30054510›Full record

ArticleScientific reports2018

Validating Antibodies for Quantitative Western Blot Measurements with Microwestern Array.

Rick J Koch, Anne Marie Barrette, Alan D Stern, Bin Hu, Mehdi Bouhaddou, Evren U Azeloglu, Ravi Iyengar, Marc R Birtwistle

Open access · goldAbstract read
In one paragraph

Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.

0numbers the graph read from it
0cells of the map it votes in
9citing papers in PubMed
0.6field-weighted citation impact, top 31% 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

9 citing papers in PubMed, 20 citations in OpenAlex.

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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

8 authors at 2 institutions in 1 country.

Rick J KochDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Anne Marie BarretteDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.ORCID 0000-0003-1452-9725
Alan D SternDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Bin HuDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Mehdi BouhaddouDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Evren U AzelogluDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.ORCID 0000-0001-6137-109X
Ravi IyengarDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. ravi.iyengar@mssm.edu.
Marc R BirtwistleDepartment of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. mbirtwi@clemson.edu.
Icahn School of Medicine at Mount Sinai · USClemson University · US

Funding

Drug Combination Signatures for Prediction and Mitigation of ToxicityU54HG008098 · NHGRI · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · PI BIRTWISTLE, MARC R., BIRTWISTLE, MARC R. · 2014 to 2019
$11.8M
Teaching biomedical and pharmacological trainees to produce FAIR data for AI & ML applicationsT32GM062754 · NIGMS · MOUNT SINAI SCHOOL OF MEDICINE OF NYU · PI SCHLESSINGER, AVNER · 2001 to 2023
$6.2M
Predicting the Heterogeneity of Cell-Fate DecisionsR01GM104184 · NIGMS · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · PI BIRTWISTLE, MARC R. · 2014 to 2018
$1.8M
NHGRI NIH HHS U54 HG008098NIGMS NIH HHS R01 GM104184NIGMS NIH HHS T32 GM062754
6 · The paper itself

Abstract

Fluorescence-based western blots are quantitative in principal, but require determining linear range for each antibody. Here, we use microwestern array to rapidly evaluate suitable conditions for quantitative western blotting, with up to 192 antibody/dilution/replicate combinations on a single standard size gel with a seven-point, two-fold lysate dilution series (~100-fold range). Pilot experiments demonstrate a high proportion of investigated antibodies (17/24) are suitable for quantitative use; however this sample of antibodies is not yet comprehensive across companies, molecular weights, and other important antibody properties, so the ubiquity of this property cannot yet be determined. In some cases microwestern struggled with higher molecular weight membrane proteins, so the technique may not be uniformly applicable to all validation tasks. Linear range for all validated antibodies is at least 8-fold, and up to two orders of magnitude. Phospho-specific and total antibodies do not have discernable trend differences in linear range or limit of detection. Total antibodies generally required higher working concentrations, but more comprehensive antibody panels are required to better establish whether this trend is general or not. Importantly, we demonstrate that results from microwestern analyses scale to normal "macro" western for a subset of antibodies.

Indexed as

AntibodiesBlotting, WesternEvaluation Studies as TopicFluorescenceHumansMembrane ProteinsAntibodiesMembrane Proteins

Identifiers

PMID30054510
PMCPMC6063895
OpenAlexW2950251081

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.