Compare the Difference Between Similar Terms

Difference Between

Home / 科学与自然 / Science / Biology / Biochemistry /Difference Between Stacking Gel and Separating Gel

Difference Between Stacking Gel and Separating Gel

February 25, 2018Posted byDr.Samanthi

Key Difference – Stacking Gel vs SeparatingGel

The terms stacking gel and separating gel are used in explaining theSDS-PAGE technique. SDS-PAGE or sodium dodecyl sulfate-polyacrylamidegel electrophoresisis a laboratory technique that is used to separate protein molecules based on theirmolecular weights. The theory behind the technique is that proteins with different molecular weights show different migration rates; low molecular weight molecules migrate faster whereas high molecular weight molecules migrate slowly. The medium for migration is a gel. There are two types of gels named as stacking gel and separating gel. Thekey differencebetween stacking gel and separating gel is thatthepHof the stacking gel is 6.8 whereas the pH of the separating gel is 8.8.

CONTENTS

1.Overview and Key Difference
2.What is Stacking Gel
3.What is Separating Gel
4.Relationship Between Stacking Gel and Separating Gel
5.Side by Side Comparison – Stacking Gel vs Separating Gel in Tabular Form
6.Summary

堆积胶是什么?

Stacking gel is a low concentrated polyacrylamide gel that is placed on the top of more concentrated resolving gel (separating gel) in SDS-PAGE technique. The stacking gel is used to improve the resolution of electrophoresis. The resolution increases because of the difference between concentrations of stacking gel and resolving gel that effect on the proteins in the sample. Since the concentration of polyacrylamide in stacking gel is low, the pore size is higher. This also helps in increasing the separation.

The stacking gel is a loose polyacrylamide gel with big pores of about 7% polyacrylamide. These pores do not act as a considerable barrier for large protein molecules. Hence this gel affects slightly on the mobility of those proteins. This makes separation according to the mobility and size of protein by concentrating them in between two gels.

Difference Between Stacking Gel and Separating Gel

Figure 01: SDS-PAGE Acrylamide Gel

The pH of the stacking gel is 6.8. Its pH isacidicthan that of resolving gel by 2 pH units. This pH implies a lower ionic strength hence a higher electrical resistance. This provokes the mobility of proteins than other charged particles present in the gel.

What is Separating Gel?

Separating gel or resolving gel of an SDS-PAGEtechnique is a highly concentrated polyacrylamide gel that is placed on the top of low concentrated stacking gel. The separating gel contains a high amount of polyacrylamide (10%). The pH of this gel is maintained as pH=8.8, which is a higher pH level than that of the stacking gel.

Key Difference Between Stacking Gel and Separating Gel

图2:凝胶电泳装置

When the protein molecules reach the separating gel, the migration of those molecules is slowed down because the separating gel is a high concentration gel with a small pore size that can act as a considerable barrier for the movement of the protein molecules. This slow down allows the other proteins are migrating slowly to catch up, resulting in a narrow, concentrated band in between the two gels.

What is the Relationship Between Stacking Gel and Separating Gel?

  • Both Stacking Gel and Separating Gel types are used in the same apparatus.

What is the Difference Between Stacking Gel and Separating Gel?

Stacking Gel vs Separating Gel

Stacking gel is a low concentrated polyacrylamide gel that is placed on the top of more concentrated resolving gel (separating gel) in SDS-PAGE technique. Separating gel or resolving gel of an SDS-PAGE technique is a highly concentrated polyacrylamide gel that is placed on the top of a low concentrated stacking gel.
Placement
Stacking gel is placed on the resolving (separating) gel. Separating gel is placed on the bottom of the container used for gel electrophoresis.
Concentration
The concentration of stacking gel is high. The concentration of separating gel is low.
Polyacrylamide Content
Stacking gel contains about 7% polyacrylamide. Separating gel contains about 10% polyacrylamide.
pH
The pH of the stacking gel is 6.8. The pH of the separating gel is 8.8.
Pore Size
Large pore sizes are present in stacking gel. Small pore sizes are present in separating gel.
Resolution
Stacking gel gives better resolution. Separating gel gives a poor resolution.

Summary –Stacking Gel vs SeparatingGel

Stacking gel and separating gel are two types of polyacrylamide gels used to get better separation of protein molecules in a given sample. The difference between stacking gel and separating gel is that the pH of the stacking gel is 6.8 whereas the pH of the separating gel is 8.8.

Reference:

1.“ Polyacrylamide Gel Electrophoresis.” Polyacrylamide Gel Electrophoresis (Theory) : Molecular Biology Virtual Lab II : Biotechnology and Biomedical Engineering : Amrita Vishwa Vidyapeetham Virtual Lab.Available here
2.“何w SDS-PAGE Works.” Bitesize Bio, 16 Feb. 2018.Available here
3.“SDS-PAGE stacking gel – BICH 608 (Polymenis-Pettigrew).” Google Sites.Available here

Image Courtesy:

1.’SDS-PAGE Acrylamide gel’By Bensaccount at English Wikipedia,(CC BY 3.0)viaCommons Wikimedia
2.’Gel electrophoresis apparatus’ By Jeffrey M. Vinocur – Own work,(CC-BY 2.5)viaCommons Wikimedia

Related posts:

Difference Between Amylopectin and GlycogenDifference Between Amylopectin and Glycogen Difference Between Fatty Acid Synthesis and Beta OxidationDifference Between Fatty Acid Synthesis and Beta Oxidation Difference Between Mode of Action and Mechanism of ActionDifference Between Mode of Action and Mechanism of Action Difference Between Respiration and Cellular RespirationDifference Between Respiration and Cellular Respiration Difference Between Visceral Fat and Subcutaneous FatDifference Between Visceral Fat and Subcutaneous Fat

Filed Under:BiochemistryTagged With:Compare Stacking Gel and Separating Gel,Separating Gel,Separating Gel Concentration,Separating Gel Definition,Separating Gel pH,Separating Gel Placement,Separating Gel Pores,Separating Gel Resolution,Separating Gel Use,Stacking Gel,Stacking Gel and Separating Gel Differences,Stacking Gel and Separating Gel Similarities,Stacking Gel Concentration,Stacking Gel Definition,Stacking Gel pH,Stacking Gel Placement,Stacking Gel Pores,Stacking Gel Resolution,Stacking Gel Use,Stacking Gel vs Separating Gel

关于the Author:Dr.Samanthi

Dr. Samanthi Udayangani holds a BSc special degree in Plant Science, a M Sc. in Molecular and Applied Microbiology and a PhD in Applied Microbiology. She has more than eight years of research experience in the fields of beneficial soil microorganisms and Biofertilizers. Her research interests are Nitrogen fixing bacteria, Phosphate solubilizing microorganisms and plant-microbe interactions. She writes articles in the subjects of Microbiology, Pathology, Diseases, Biochemistry, Molecular Biology and Botany. You can read her researches onResearchGate

Leave a ReplyCancel reply

Your email address will not be published.Required fields are marked*

Request Article

Featured Posts

Difference Between Coronavirus and Cold Symptoms

Difference Between Coronavirus and Cold Symptoms

Difference Between Coronavirus and SARS

Difference Between Coronavirus and SARS

Difference Between Coronavirus and Influenza

Difference Between Coronavirus and Influenza

Difference Between Coronavirus and Covid 19

Difference Between Coronavirus and Covid 19

You May Like

Difference Between Bratwurst and Sausage and Salami

Difference Between Rubbing Alcohol and Hand Sanitizer

Difference Between Rubbing Alcohol and Hand Sanitizer

Difference Between iPhone 4S and BlackBerry Torch 9860

Difference Between Pampers and Huggies

Difference Between Pampers and Huggies

Difference Between Absorptivity and Molar Absorptivity

Difference Between Absorptivity and Molar Absorptivity

Latest Posts

  • What is the Difference Between Intrahepatic and Extrahepatic Cholestasis
  • What is the Difference Between Aerobic and Anaerobic Biodegradation
  • What is the Difference Between Blue and Lilac French Bulldog
  • What is the Difference Between Beta Thalassemia Major and Minor
  • What is the Difference Between Creutzfeldt Jakob Disease and Mad Cow Disease
  • What is the Difference Between Leukorrhea and Steatorrhea
  • Home
  • Vacancies
  • 关于
  • Request Article
  • Contact Us

Copyright © 2010-2018Difference Between. All rights reserved.Terms of Useand Privacy Policy:Legal.