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Related Concept Videos

Western Blotting01:15

Western Blotting

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.
Southern Blot02:57

Southern Blot

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...

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Related Experiment Video

Updated: Jun 23, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

"Rainbow" western blotting.

Stan Krajewski1

  • 1Burnham Institute for Medical Research, 10901 N. Torrey Pines Rd, La Jolla, CA, 92037, USA. stan@burnham.org

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed
Summary
This summary is machine-generated.

The Rainbow western method detects multiple proteins on one blot using horseradish peroxidase (HRPase) and different colored substrates. This technique streamlines analysis of protein expression and modifications in biological samples.

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Related Experiment Videos

Last Updated: Jun 23, 2026

Western Blotting: Sample Preparation to Detection
07:45

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology
05:59

Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology

Published on: September 26, 2020

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

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunodetection

Background:

  • Western blotting is a key technique for protein analysis.
  • Detecting multiple proteins typically requires separate blots or complex protocols.
  • Analyzing protein expression and modifications often involves multiple targets.

Purpose of the Study:

  • To introduce a novel method for multiplexed antigen detection on a single protein blot.
  • To enable simultaneous visualization of up to four different antigens with distinct colors.
  • To provide a consolidated approach for analyzing protein expression and post-translational modifications.

Main Methods:

  • Utilizes horseradish peroxidase (HRPase)-based detection.
  • Employs sequential application of four distinct HRPase-colorimetric substrates (black, brown, red, green).
  • Allows for simultaneous display of multiple antigens on a single membrane.

Main Results:

  • Successful detection of four different antigens on the same blot.
  • Distinct colorimetric signals (black, brown, red, green) for each antigen.
  • Demonstrates the feasibility of multiplexed protein detection.

Conclusions:

  • The Rainbow western method offers a consolidated approach for protein analysis.
  • This technique is valuable for studying signaling pathways and protein modifications.
  • It is particularly useful for analyzing comigrating proteins and isoforms.