title: Dealing with large sample sizes: comparison of a new one spot dot blot method to western blot.
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.
authors: Sulistyo Emantoko Dwi
authors: SE
authors: Putra
authors: Oleg
authors: O
authors: Tsuprykov
authors: Karoline
authors: K
authors: Von Websky
authors: Teresa
authors: T
authors: Ritter
authors: Christoph
authors: C
authors: Reichetzeder
authors: Berthold
authors: B
authors: Hocher
mesh_terms: D000469:Alkaline Phosphatase; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D024042:Collagen Type I / Q000378:metabolism; D004195:Disease Models, Animal; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006735:Horseradish Peroxidase; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007668:Kidney / Q000378:metabolism*; D007674:Kidney Diseases / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D009392:Nephrectomy; D017208:Rats, Wistar; D015203:Reproducibility of Results; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D013048:Specimen Handling
chemical_list: D015415:Biomarkers; D024042:Collagen Type I; D008565:Membrane Proteins; D051899:Smad2 Protein; D051900:Smad3 Protein; C113267:nephrin; D006735:Horseradish Peroxidase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000469:Alkaline Phosphatase