Evidence map›Paper›PMID 20926088›Full record

ArticleJournal of chromatography. A2010

Repeated probing of Southwestern blots using alkaline phosphatase stripping.

Yinshan Jia, Daifeng Jiang, Harry W Jarrett

Abstract read
In one paragraph

Article in Journal of chromatography. A, 2010. 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 46% 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. Southwestern Blotting Assay.Methods in molecular biology (Clifton, N.J.) · 2015
    Article
  2. DNA-protein interactions: methods for detection and analysis.Molecular and cellular biochemistry · 2012
    Review
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

3 authors at 1 institution in 1 country.

Yinshan JiaDepartment of Chemistry, University of Texas San Antonio, San Antonio, TX 78249, USA.
Daifeng Jiang
Harry W Jarrett
The University of Texas at San Antonio · US

Funding

POLYNUCLEOTIDE HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHYR01GM043609 · NIGMS · UNIVERSITY OF TEXAS SAN ANTONIO · PI JARRETT, HARRY W · 1989 to 2013
$3.1M
NIGMS NIH HHS 5R01GM043609NIGMS NIH HHS R01 GM043609
6 · The paper itself

Abstract

Southwestern blotting is when a DNA sequence is used to probe DNA-binding proteins on an electrophoretic gel blot. It would be highly desirable to be able to probe a blot repeatedly with different DNA sequences. Alkaline phosphatase can remove 5'-phosphoryl groups from DNA and radiolabeled 5'-(32)P-DNA probes are commonly used in Southwestern blotting. Here is shown that once probed, the radioisotope signal on the blot can be effectively removed by brief digestion with alkaline phosphatase, and the blot can then be repeatedly probed at least six times with different DNA probes. This exceeds the repetitions possible with another commonly used method using SDS. The technique can be used with either one-dimensional or multi-dimensional Southwestern blots and does not have a large effect on the phosphorylation state of the blotted proteins. An alternative method using T4 polynucleotide kinase stripping is also introduced but was less well characterized.

Indexed as

Alkaline PhosphataseAnimalsBlotting, SouthwesternCattleCell NucleusDNADNA-Binding ProteinsEquipment ReuseFluorocarbon PolymersHEK293 CellsHumansOligonucleotidesPhosphorus IsotopesPhosphorylationPolyvinylsSodium Dodecyl SulfateAlkaline PhosphataseDNADNA-Binding ProteinsFluorocarbon PolymersOligonucleotidesPhosphorus Isotopespolyvinylidene fluoridePolyvinylsSodium Dodecyl SulfateTranscription Factors

Identifiers

PMID20926088
PMCPMC2964665
OpenAlexW2081142928

What OpenQuestion holds

Textmetadata
LicenceTDM
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.