Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Western Blotting01:15

Western Blotting

20.8K
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
20.8K
Reliability and Validity01:29

Reliability and Validity

14.1K
Reliability and validity are two important considerations that must be made with any type of data collection. Reliability refers to the ability to consistently produce a given result. In the context of psychological research, this would mean that any instruments or tools used to collect data do so in consistent, reproducible ways.
14.1K
Southern Blot02:57

Southern Blot

23.1K
Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
23.1K
Data Validation01:15

Data Validation

2.0K
Method validation is a crucial process in analytical chemistry designed to confirm that a given method consistently produces reliable and high-quality results. This process is essential when a method is applied to different sample matrices or when procedural modifications are made, ensuring that the results meet acceptable standards across various applications.
Key parameters for method validation include:
2.0K
Data Validation01:03

Data Validation

6.8K
Data validation is an essential part of a comprehensive assessment. Validation is confirming or verifying and opening the door to gathering more assessment data as it clarifies vague or unclear data. The process of checking and verifying the collected information is called data validation. The primary purpose of data validation is to ensure data is as free from error, bias, and misinterpretation as possible.
Nursing assessment guides are generally based on holistic models rather than medical...
6.8K
Antibody Structure01:10

Antibody Structure

65.6K
Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
65.6K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Author Correction: LILRB3 genetic variation is associated with kidney transplant failure in African American recipients.

Nature medicine·2025
Same author

LILRB3 genetic variation is associated with kidney transplant failure in African American recipients.

Nature medicine·2025
Same author

Multiscale mapping of transcriptomic signatures for cardiotoxic drugs.

Nature communications·2024
Same author

A high-throughput microfabricated platform for rapid quantification of metastatic potential.

Science advances·2024
Same author

Low-frequency ERK and Akt activity dynamics are predictive of stochastic cell division events.

NPJ systems biology and applications·2024
Same author

Integrated multiomics implicates dysregulation of ECM and cell adhesion pathways as drivers of severe COVID-associated kidney injury.

medRxiv : the preprint server for health sciences·2024

Related Experiment Video

Updated: Feb 7, 2026

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies
11:01

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

52.5K

Validating Antibodies for Quantitative Western Blot Measurements with Microwestern Array.

Rick J Koch1, Anne Marie Barrette1, Alan D Stern1

  • 1Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.

Scientific Reports
|July 29, 2018
PubMed
Summary
This summary is machine-generated.

The microwestern array rapidly determines optimal conditions for quantitative western blotting. This method validates antibody performance, ensuring reliable results for western blot assays.

More Related Videos

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis
11:43

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

28.0K
Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting
08:13

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Published on: April 9, 2019

14.9K

Related Experiment Videos

Last Updated: Feb 7, 2026

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies
11:01

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

52.5K
The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis
11:43

The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

Published on: February 5, 2014

28.0K
Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting
08:13

Robust Comparison of Protein Levels Across Tissues and Throughout Development Using Standardized Quantitative Western Blotting

Published on: April 9, 2019

14.9K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • Quantitative western blotting is crucial for accurate protein analysis.
  • Determining the linear range for each antibody is essential but time-consuming.
  • Antibody validation is critical for reproducible scientific research.

Purpose of the Study:

  • To introduce and validate the microwestern array as a rapid method for optimizing quantitative western blotting conditions.
  • To assess the suitability of various antibodies for quantitative western blot analysis.
  • To evaluate the linear range and performance characteristics of antibodies using the microwestern array technique.

Main Methods:

  • Utilized microwestern arrays to test up to 192 antibody/dilution/replicate combinations simultaneously.
  • Employed a seven-point, two-fold lysate dilution series (~100-fold range) for comprehensive analysis.
  • Compared microwestern array results with traditional "macro" western blot methods.

Main Results:

  • A high proportion of tested antibodies (17/24) were found suitable for quantitative western blotting.
  • Validated antibodies demonstrated a linear range of at least 8-fold, up to two orders of magnitude.
  • Microwestern array results successfully scaled to conventional western blot outcomes for a subset of antibodies.
  • No discernible trend differences were observed between phospho-specific and total antibodies regarding linear range or limit of detection.

Conclusions:

  • The microwestern array is an effective tool for rapidly evaluating antibody suitability and optimizing conditions for quantitative western blotting.
  • The technique provides reliable data that correlates with traditional western blot methods.
  • Further studies with broader antibody panels are needed to confirm general applicability, especially for high molecular weight proteins.