Lectin Blotting
Lectin Blotting - Related Videos
09:25
Lectin-based Isolation and Culture of Mouse Embryonic Motoneurons
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Cited by 21 •
2011
An alternative way of isolating mouse embryonic motoneurons from the spinal cord is described. The method takes into account the fact that lectin can bind to the low affinity nerve growth factor receptor p75NTR. This lectin-based preplating allows a purification similar to that with a specific antibody against the p75NTR.
20:23
A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
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Cited by 13 •
2009
Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.
08:11
Measuring Influenza Neuraminidase Inhibition Antibody Titers by Enzyme-linked Lectin Assay
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Cited by 54 •
2016
We describe the enzyme-linked lectin assay (ELLA) for measuring influenza neuraminidase (NA)-inhibition antibody titers in sera. The assay uses peanut agglutinin to quantify galactose residues that become accessible when NA removes sialic acid from fetuin-coated, 96-well plates.
08:48
The Western Blot
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2023
Western Blotting is used to identify the presence of specific proteins in electrophoretically separated samples. Following separation by a technique known as sodium dodecyl sulfate polyacrylamide gel electrophoresis, or SDS-PAGE, western transfer is used to move proteins from a polyacrylamide gel onto a piece of membrane which traps the proteins in their respective locations. Next, the membranes are probed with antibodies in a process called immunboblotting. Immunoblotting uses...
01:15
Western Blotting
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2023
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.
FAQs
Lectin choice determines which carbohydrate features become visible. Because individual lectins recognize specific glycan motifs, changing the labeled lectin can reveal different subsets of glycoproteins in the same sample. The resulting pattern reflects the interaction between the probe and available carbohydrate structures, allowing researchers to examine differences in glycosylation rather than treating all glycoproteins as equivalent.
Gel electrophoresis separates the proteins before detection, while membrane transfer places those separated components on a surface that can be exposed to the labeled lectin. This arrangement connects each signal to a particular position within the separated protein pattern. As a result, researchers can compare which protein regions contain recognized carbohydrate structures instead of observing an undifferentiated mixture.
The two approaches emphasize different molecular information. Antibody-based Western blotting primarily asks whether a particular protein identity is present, whereas lectin-based detection focuses on carbohydrate structures attached to proteins. Lectin Blotting therefore complements antibody analysis by revealing glycosylation-related differences that may not be apparent when samples are evaluated through protein identity alone.
The workflow begins by separating proteins through gel electrophoresis and transferring them to a membrane. The membrane is then exposed to a labeled lectin selected for its carbohydrate-binding properties. Detection of the resulting signal shows which separated protein components contain the carbohydrate features recognized by that lectin, linking sample preparation to glycoprotein pattern analysis.
Researchers can examine the lectin-detected patterns produced by different samples and identify changes in the presence or distribution of recognized carbohydrate structures. Using the same separation and lectin-detection approach makes those patterns comparable. Differences can provide evidence of altered glycosylation or protein processing, helping distinguish biochemical states associated with different cellular conditions.
This technique can investigate how carbohydrate structures relate to protein modification and function, including changes in cellular processing. It is also useful for examining glycosylation alterations associated with disease-related states. Because the readout concerns recognized carbohydrate motifs, the method adds structural context to studies of glycoproteins beyond what protein identity alone can provide.