Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Western Blotting

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 Blot

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...

You might also read

Related Articles

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

Sort by

Same author

The role of DNMT3B, GSK-3β and GJA3 in lens epithelial cell apoptosis and age-related cataract.

BMC ophthalmology·2026

Same author

The dynamics of glycosylation in trophoblast cells: development, function, and pregnancy-related disorders.

Annals of medicine·2026

Same author

Integrated Proteomics and Metabolomics Analysis in Hippocampus of Rats With T2DM-Associated Cognitive Decline.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology·2026

Same author

Deficient extravillous trophoblast invasion caused by impaired sialylation-Siglec-7 interaction contributes to recurrent pregnancy loss.

Cell death & disease·2026

Same author

DNA Methylation Markers for Pregnancy Hypertension via Machine Learning Methods.

Hypertension (Dallas, Tex. : 1979)·2026

Same author

Tracheobronchopathia osteochondroplastica: A rare case report.

Medicine·2026

Same journal

Correction.

Andrologia·2023

Same journal

The combination of ginger and zinc supplement could improve lead-induced reproductive dysfunction by inhibiting apoptosis mediated by oxidative damage and inflammation.

Andrologia·2022

Same journal

Testosterone and lifespan in males: At the right time and the right level.

Andrologia·2022

Same journal

CORRIGENDUM.

Andrologia·2022

Same journal

CORRIGENDUM.

Andrologia·2022

Same journal

Microfluidic devices employing chemo- and thermotaxis for sperm selection can improve sperm parameters and function in patients with high DNA fragmentation.

Andrologia·2022

See all related articles

Search research articles

Related Experiment Video

Updated: Oct 21, 2025

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

Identifying new sperm Western blot loading controls.

Ying Feng¹, Ruohan Wang^2,3, Dongmei Su^2,3

¹West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, China.

|September 3, 2021

Summary

This summary is machine-generated.

Identifying stable housekeeping proteins is crucial for accurate sperm protein analysis. This study found β-tubulin, Cullin-1 (CUL1), and F-box only protein 7 (FBXO7) to be reliable internal controls for Western Blot experiments in sperm.

Keywords:

cellular location housekeeping protein preservation sperm transcriptome

More Related Videos

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Western Blotting: Sample Preparation to Detection

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Related Experiment Videos

Last Updated: Oct 21, 2025

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

Published on: December 30, 2013

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

Published on: November 20, 2014

Western Blotting: Sample Preparation to Detection

Western Blotting: Sample Preparation to Detection

Published on: October 14, 2010

Area of Science:

Biochemistry
Molecular Biology
Reproductive Biology

Background:

Accurate protein expression analysis is vital in biological and medical research.
Housekeeping proteins serve as essential loading controls for normalizing protein expression.
Widely used normalization proteins exhibit variable expression in sperm samples, compromising data standardization.

Purpose of the Study:

To identify stably expressed proteins in human sperm suitable for use as housekeeping controls.
To evaluate the suitability of classical housekeeping proteins and novel candidates for sperm protein normalization.

Main Methods:

Analysis of published sperm transcriptome data to identify candidate proteins with stable expression (coefficient of variation < 0.35).
Validation of selected proteins using quantitative real-time polymerase chain reaction (qRT-PCR), Western Blot (WB), and immunocytochemistry.
Assessment of protein expression stability across sperm samples from 85 individuals.

Main Results:

Classical housekeeping proteins like glyceraldehyde 3-phosphate dehydrogenase, actin, and histone H3 showed significant variability in sperm.
Only β-tubulin demonstrated consistent expression among classical housekeeping proteins in sperm samples.
Cullin-1 (CUL1) and F-box only protein 7 (FBXO7) exhibited stable expression and were identified as suitable internal controls for WB in sperm.

Conclusions:

Standard normalization proteins are inadequate for accurate sperm protein quantification.
β-tubulin, CUL1, and FBXO7 are recommended as reliable housekeeping proteins for Western Blot analysis in sperm studies.
CUL1 and FBXO7 are particularly suggested as housekeeping proteins for total protein analysis in sperm, considering their stable expression and cellular localization.