Cell Surface Protein Isolation
Cell Surface Protein Isolation - Related Videos
04:58
Isolation of Cell-Surface and Intracellular Proteins from an Astrocyte Culture
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2025
This video demonstrates the isolation of cell-surface and intracellular proteins from an astrocyte culture. The astrocyte culture is placed on ice to inhibit endocytosis, and then the cell-surface proteins are labeled with a biotinylation reagent. The cells are lysed to release the biotinylated cell-surface proteins and non-biotinylated intracellular proteins. Finally, streptavidin-coated beads are used to separate the biotinylated cell-surface proteins from the intracellular proteins.
09:12
Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins
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Cited by 4 •
2019
Here we present a protocol for murine in vivo labeling of glomerular cell surface proteins with biotin. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of the protein of interest.
12:03
Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells
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Cited by 3 •
2017
A modified density centrifugation gradient-based methodology was utilized to isolate epithelial cells from Rhipicephalus microplus gut tissue. Surface-bound proteins were biotinylated and purified through streptavidin magnetic beads for utilization in downstream applications.
03:33
Cation Exchange Chromatography: A Technique to Isolate Target Recombinant Protein from Insect Cell Lysate Based on Net Surface Charge
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2025
This video demonstrates the cation exchange chromatography technique to isolate recombinant protein from the insect cell protein lysate.
10:40
A Method of Targeted Cell Isolation via Glass Surface Functionalization
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Cited by 6 •
2016
This protocol describes customizable surface functionalization of the desthiobiotin, streptavidin, and APTES system in order to isolate specific cell types of interest. In addition, this manuscript covers the applications, optimization, and verification of this process.
FAQs
Selectivity comes from using a membrane-impermeant biotinylation reagent, which labels accessible protein domains outside the cell while limiting labeling of intracellular proteins. This chemical distinction creates a surface-enriched population rather than a general cellular protein mixture. Maintaining controlled labeling conditions is therefore central to interpreting whether a detected protein represents exterior exposure.
Streptavidin-coated beads selectively retain the biotin-labeled proteins after the cell-surface labeling step. Washing removes material that has not been specifically captured, while elution releases the enriched proteins for downstream analysis. This sequence separates affinity capture from measurement and helps produce a profile focused on labeled surface-associated material rather than the full cellular protein pool.
An enriched surface profile can show whether receptor abundance changes and can support studies of membrane trafficking, cell adhesion, and signaling. These measurements focus attention on proteins exposed to the cell’s environment, where altered abundance or localization can affect communication. The approach therefore connects biochemical enrichment with functional questions about cellular interactions.
Immunoblotting and mass spectrometry provide complementary ways to examine the isolated fraction. Immunoblotting can assess selected proteins, whereas mass spectrometry can characterize proteins represented in the enriched sample more broadly. Choosing between them depends on whether the experiment emphasizes particular targets or a wider surface-protein profile.
The practical sequence begins with controlled labeling of extracellular protein domains, followed by collection of the labeled material with streptavidin-coated beads. Researchers then wash the beads to remove nonspecifically retained proteins and elute the captured fraction. The recovered sample can subsequently undergo immunoblotting or mass spectrometry to evaluate its surface-protein composition.
Cell Surface Protein Isolation is especially useful when the question concerns changes at the cell boundary rather than total cellular protein content. Researchers can apply it to investigate receptor abundance, membrane trafficking, cell adhesion, or signaling, and to compare surface profiles during development, disease, or treatment. Its value lies in linking external protein exposure with changing biological states.
In biology, the method helps clarify how cells communicate with their environment by focusing analysis on proteins exposed outside the cell. Surface-protein profiles can reveal changes associated with development, disease, or treatment, providing biochemical evidence for altered cellular interactions. These results may also help distinguish a change in surface presentation from the broader presence of proteins inside the cell.