Polyacrylamide Gel Electrophoresis
Polyacrylamide Gel Electrophoresis - Related Videos
08:44
Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)
0 Views •
Cited by 104 •
2009
Denaturing urea polyacrylamide gel electrophoresis is used to separate single-stranded DNA or RNA up to a limit of 500 nucleotides. Urea in combination with heat denatures samples and unstructured single strands migrate within the gel matrix according to their molecular weight.
08:55
Characterization of Multi-subunit Protein Complexes of Human MxA Using Non-denaturing Polyacrylamide Gel-electrophoresis
0 Views •
Cited by 3 •
2016
This article describes a simple and rapid protocol to evaluate the oligomeric state of the dynamin-like GTPase MxA protein from lysates of human cells using a combination of non-denaturing PAGE with western blot analysis.
07:31
Two-Dimensional Gel Electrophoresis
0 Views •
2023
Two-dimensional gel electrophoresis (2DGE) is a technique that can resolve thousands of biomolecules from a mixture. This technique involves two distinct separation methods that have been coupled together: isoelectric focusing (IEF) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). This physically separates compounds across two axes of a gel by their isoelectric points (an electrochemical property) and their molecular weights. The procedure in this video covers the main...
04:15
Polyacrylamide Gel Electrophoresis: An Analytical Technique to Detect Heparan Sulfates of Varying Chain Lengths Isolated from Mouse Lung Tissue
0 Views •
2025
This video demonstrates the separation procedure of heparan sulfates isolated from mouse lung tissue using polyacrylamide gel electrophoresis.
12:03
Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates
0 Views •
Cited by 92 •
2011
In this video, we describe the characterization of multiprotein complexes (MPCs) by blue native polyacrylamide gel electrophoresis (BN-PAGE). In a first dimension, dialyzed cellular lysates are separated by BN-PAGE to identify individual MPCs. In a second dimension SDS-PAGE, MPCs of interest are further subdivided to analyze their constituents by immunoblotting.
FAQs
In SDS-PAGE, sodium dodecyl sulfate makes separation depend primarily on molecular size, allowing protein components to be compared by their relative migration. Native PAGE preserves information associated with charge, shape, and protein complexes. Choosing between these formats determines whether the experiment emphasizes size-based comparison or the state of proteins in assembled forms.
The polyacrylamide matrix provides a porous environment through which charged biomolecules migrate under an electric field. Its presence enables proteins to become spatially separated rather than remaining as a combined sample. This separation is essential for comparing components individually, assessing protein purity, and preparing samples for later antibody-based analysis.
Native PAGE retains features that denaturing SDS-PAGE does not prioritize, including protein charge, shape, and association into complexes. Consequently, the resulting separation can provide information about proteins in assembled forms rather than only their size-related behavior. This distinction is relevant when infection or immune studies examine protein organization or complex formation.
PAGE can be performed before immunoblotting to resolve proteins within a complex sample. The separated components then provide an organized basis for antibody-based detection and comparison. In immunology and infection research, this workflow supports examination of antigen expression, immune recognition, and infection-associated changes by linking antibody reactivity to separated protein components.
Under denaturing SDS-PAGE conditions, PAGE supports assessment of apparent molecular weight through size-based separation. It can also help evaluate protein purity by showing whether a sample contains a limited or varied set of resolved components. These outcomes make the technique useful for characterizing preparations before additional immunological or infection-related analyses.
PAGE helps resolve proteins originating from pathogens or their hosts, allowing researchers to compare protein composition and infection-associated changes. In immunology, the separated material can support studies of antigen expression and immune recognition. Its value comes from connecting protein separation with questions about how infection alters host or pathogen-associated protein patterns.