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Dealing with large sample sizes: comparison of a new one spot dot blot method to western blot
- Publication Year :
- 2014
-
Abstract
- Background: Western blot is the gold standard method to determine individual protein expression levels. However, western blot is technically difficult to perform in large sample sizes because it is a time consuming and labor intensive process. Dot blot is often used instead when dealing with large sample sizes, but the main disadvantage of the existing dot blot techniques, is the absence of signal normalization to a housekeeping protein. Methods: In this study we established a one dot two development signals (ODTDS) dot blot method employing two different signal development systems. The first signal from the protein of interest was detected by horseradish peroxidase (HRP). The second signal, detecting the housekeeping protein, was obtained by using alkaline phosphatase (AP). Results: Inter-assay results variations within ODTDS dot blot and western blot and intra-assay variations between both methods were low (1.04 - 5.71%) as assessed by coefficient of variation. Conclusions: ODTDS dot blot technique can be used instead of western blot when dealing with large sample sizes without a reduction in results accuracy.
- Subjects :
- Male
Coefficient of variation
Blotting, Western
Immunoblotting
Dot blot
Smad2 Protein
Kidney
Horseradish peroxidase
Nephrectomy
General Biochemistry, Genetics and Molecular Biology
Collagen Type I
Specimen Handling
Western blot
medicine
Animals
Humans
Smad3 Protein
Rats, Wistar
Horseradish Peroxidase
biology
medicine.diagnostic_test
Northwestern blot
Kidney metabolism
Glyceraldehyde-3-Phosphate Dehydrogenases
Membrane Proteins
Reproducibility of Results
Reverse northern blot
Alkaline Phosphatase
Molecular biology
Blot
Disease Models, Animal
biology.protein
Kidney Diseases
Institut für Ernährungswissenschaft
Biomarkers
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....b7d80f441325ed6fe105e59e28cd2efa