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Molecular Profiling

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Cover of 'Molecular Profiling'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 Tumor staging and grading: a primer.
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    Chapter 2 Clinical trial design in the age of molecular profiling.
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    Chapter 3 Personalized medicine: ethics for clinical trials.
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    Chapter 4 Reduction of preanalytical variability in specimen procurement for molecular profiling.
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    Chapter 5 The human side of cancer biobanking.
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    Chapter 6 Introduction to genomics.
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    Chapter 7 Genomic profiling: cDNA arrays and oligoarrays.
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    Chapter 8 Genome-wide methylation profiling in archival formalin-fixed paraffin-embedded tissue samples.
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    Chapter 9 An Overview of MicroRNA Methods: Expression Profiling and Target Identification.
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    Chapter 10 Antibody Validation by Western Blotting
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    Chapter 11 Laser Capture Microdissection: Arcturus(XT) Infrared Capture and UV Cutting Methods.
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    Chapter 12 Antibody microarrays: analysis of cystic fibrosis.
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    Chapter 13 Tissue microarrays as a tool in the discovery and validation of predictive biomarkers.
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    Chapter 14 Reverse-phase protein microarrays.
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    Chapter 15 Serum low-molecular-weight protein fractionation for biomarker discovery.
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    Chapter 16 Mass spectrometry-based biomarker discovery.
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    Chapter 17 Mitochondrial proteome: toward the detection and profiling of disease associated alterations.
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    Chapter 18 Adult neural stem cells: isolation and propagation.
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    Chapter 19 Evanescent-wave field imaging: an introduction to total internal reflection fluorescence microscopy.
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    Chapter 20 Construction and hyperspectral imaging of quantum dot lysate arrays.
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    Chapter 21 Microarray data analysis: comparing two population means.
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    Chapter 22 Bioinformatics/Biostatistics: microarray analysis.
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    Chapter 23 Structure-based functional design of drugs: from target to lead compound.
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    Chapter 24 Personalized medicine: changing the paradigm of drug development.
  26. Altmetric Badge
    Chapter 25 Grant writing tips for translational research.
  27. Altmetric Badge
    Chapter 26 Inventions and patents: a practical tutorial.
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    Chapter 27 Regulatory approval pathways for molecular diagnostic technology.
  29. Altmetric Badge
    Chapter 28 Small business development for molecular diagnostics.
Attention for Chapter 10: Antibody Validation by Western Blotting
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Chapter title
Antibody Validation by Western Blotting
Chapter number 10
Book title
Molecular Profiling
Published in
Methods in molecular biology, October 2011
DOI 10.1007/978-1-60327-216-2_10
Pubmed ID
Book ISBNs
978-1-60327-215-5, 978-1-60327-216-2
Authors

K. Alex Reeder, Michele Signore

Editors

Virginia Espina, Lance A. Liotta

Abstract

Validation of antibodies is an integral part of translational research, particularly for biomarker discovery. Validation is essential to show the specificity of the reagent (antibody) and to confirm the identity of the protein biomarker, prior to implementing the biomarker in clinical studies.Antibody validation is the procedure in which a single antibody is thoroughly assayed for sensitivity and specificity. Although a plethora of commercial antibodies exist, antibody specificity must be thoroughly demonstrated using a complex biological sample, rather than a recombinant protein, prior to use in clinical translational research. In the simplest iteration, antibody specificity is determined by the presence of a single band in a complex biological sample, at the expected molecular weight, on a western blot.Numerous western blotting procedures are available, spanning the spectrum of single blots to multiplex blots, with images and quantitation generated by manual or automated systems. The basic principles of western blotting are (a) separation of protein mixtures by gel electrophoresis, (b) transfer of the proteins to a blot, (c) probing the blot for a protein or proteins of interest, and (d) subsequent detection of the protein by chemiluminescent, fluorescent, or colorimetric methods. This chapter focuses on the chemiluminescent detection of proteins using a manual western blotting system and a vacuum-enhanced detection system (SNAP i.d.™, Millipore).

Mendeley readers

The data shown below were compiled from readership statistics for 20 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Mexico 1 5%
Unknown 19 95%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 4 20%
Student > Bachelor 4 20%
Student > Doctoral Student 4 20%
Student > Master 2 10%
Professor 1 5%
Other 3 15%
Unknown 2 10%
Readers by discipline Count As %
Agricultural and Biological Sciences 6 30%
Medicine and Dentistry 5 25%
Biochemistry, Genetics and Molecular Biology 4 20%
Immunology and Microbiology 1 5%
Veterinary Science and Veterinary Medicine 1 5%
Other 0 0%
Unknown 3 15%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 09 February 2013.
All research outputs
#2,900,237
of 3,629,816 outputs
Outputs from Methods in molecular biology
#1,438
of 2,357 outputs
Outputs of similar age
#225,361
of 282,051 outputs
Outputs of similar age from Methods in molecular biology
#85
of 177 outputs
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We're also able to compare this research output to 177 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.