Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis

After separation of whole proteins of chick neural retina by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), a number of glycoproteins can be detected by staining the gels with 125I-labeled wheat germ agglutinin (WGA) and other lectins. The glycoprotein patterns show both quantitative and qualitative changes between days 7 and 13 of development. Some of these glycoproteins can be separated by chromatography on columns of insolubilized lectins. These observations suggest that purification of some of these glycoproteins identified by staining with radioactive lectins would yield retinal antigens which may be specific for developmental stage and cell type.

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Selective Staining of Platelet Glycoproteins Using Two-Dimensional O’Farrell Gel Electrophoresis and Avidin-Biotin-Conjugated Lectins

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646036 ◽

1987 ◽

Vol 58 (04) ◽

pp. 960-963 ◽

Cited By ~ 6

Author(s):

Beate Kehrel ◽

Renate Kokott ◽

Leopold Balleisen ◽

Werner Stenzinger ◽

Kenneth J Clemetson ◽

...

Keyword(s):

Gel Electrophoresis ◽

Wheat Germ ◽

Lens Culinaris ◽

Normal Subjects ◽

Resolution Method ◽

Time Saving ◽

Selective Staining ◽

Abrus Precatorius ◽

Sds Page ◽

High Resolution Method

SummaryA time-saving and sensitive high resolution method lor the analysis and identification of platelet glycoproteins using non-radioactive compounds has been developed. Platelet proteins (50 μg) of normal subjects were separated by isoelectric focusing and SDS-PAGE. Proteins were either stained with silver or electroblotted onto nitrocellulose. Nitrocellulose blots were treated with the following biotinylated lectins : Abrus precatorius lectin, concanavalin A, Lens culinaris lectin, or wheat germ agglutinin. Staining was achieved by avidin-biotin-coupled peroxidase using 4-chloro-l-naphthol as the substrate. A rapid overview of platelet glycoproteins may be obtained by applying all the lectins to a single blot.

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Identification of Subpopulations of Human Urinary Plasminogen Activators

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661061 ◽

1984 ◽

Vol 51 (02) ◽

pp. 212-216 ◽

Cited By ~ 13

Author(s):

David A Hart ◽

Robert Kramer ◽

Witold Cieplak

Keyword(s):

Affinity Chromatography ◽

Concanavalin A ◽

Human Urine ◽

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Plasminogen Activators ◽

Lentil Lectin ◽

Sds Page ◽

Commercial Sources ◽

Individual Human

SummaryEnzymes functioning as plasminogen activators in commercial urokinase preparations and individual human urine concentrates were subjected to affinity chromatography on columns of lentil lectin-sepharose, ricin-sepharose, wheat germ agglutinin-sephar-ose, lotus lectin-sepharose and concanavalin A-sepharose.Chromatography of the enzymes from both sources yielded similar results for all lectins except lentil lectin. Urokinase from several commercial sources was approximately 50% adherent to lentil lectin-sepharose while only 5-10% of the urinary plasminogen activators from individuals was adherent to this lectin. SDS-PAGE followed by zymography indicated that the observed differences between commercial and individual samples could be due to the presence in urine concentrates of subpopulations of plasminogen activators which were absent from the commercial samples.

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Partial purification of dopamine D2 receptors using lectin affinity columns

Bioscience Reports ◽

10.1007/bf01116901 ◽

1985 ◽

Vol 5 (4) ◽

pp. 303-308 ◽

Cited By ~ 11

Author(s):

W. Mark Abbott ◽

Philip G. Strange

Keyword(s):

Sodium Chloride ◽

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Affinity Purification ◽

D2 Receptors ◽

Dopamine D2 Receptors ◽

Purification Procedure ◽

D2 Dopamine Receptor ◽

Lectin Affinity ◽

Dopamine D2

Dopamine D2 receptors, detected by [3H]spiperone Dopamine D2 receptors, detected by [3H]spiperone binding, were solubilized from bovine caudate nucleus by cholate/sodium chloride and were found to bind to wheat germ agglutinin immobilized on agarose. Specific elution could be achieved with N-acetylglucosamine whereas other sugars tested were inactive in this regard. The eluted preparation was enriched in solubilized receptors about sevenfold. The pharmaco-logical properties of the preparation were essentially unchanged by the lectin affinity purification procedure. The D2 dopamine receptor is therefore a glycoprotein. binding, were solubilized from bovine caudate nucJeus by cholate/sodium chloride and were found to bind to wheat germ agglutinin immobilized on agarose. Specific elution could be achieved with N-acetylglucosamine whereas other sugars tested were inactive in this regard. The eluted preparation was enriched in solubilized receptors about sevenfold. The pharmacological properties of the preparation were essentially unchanged by the lectin affinity purification procedure. The D2 dopamine receptor is therefore a glycoprotein.

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A novel method for live imaging of human airway cilia using wheat germ agglutinin

Scientific Reports ◽

10.1038/s41598-020-71049-z ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ryosuke Nakamura ◽

Tatsuya Katsuno ◽

Yo Kishimoto ◽

Shinji Kaba ◽

Masayoshi Yoshimatsu ◽

...

Keyword(s):

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Live Imaging ◽

Human Airway ◽

Novel Method ◽

Airway Cilia

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Serial lectin affinity chromatography with concanavalin A and wheat germ agglutinin demonstrates altered asparagine-linked sugar-chain structures of prostatic acid phosphatase in human prostate carcinoma

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/s0378-4347(97)00186-2 ◽

1997 ◽

Vol 695 (2) ◽

pp. 439-443 ◽

Cited By ~ 25

Author(s):

Ken-ichiro Yoshida ◽

Mikihiko Honda ◽

Kyoko Arai ◽

Yoshakatsu Hosoya ◽

Hideo Moriguchi ◽

...

Keyword(s):

Acid Phosphatase ◽

Concanavalin A ◽

Prostate Carcinoma ◽

Wheat Germ Agglutinin ◽

Wheat Germ ◽

Prostatic Acid Phosphatase ◽

Lectin Affinity Chromatography ◽

Sugar Chain ◽

Lectin Affinity ◽

Human Prostate Carcinoma

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Detection of Citrus Leaf and Seed Glycoproteins using Biotinylated Lectin Probes

HortScience ◽

10.21273/hortsci.26.7.910 ◽

1991 ◽

Vol 26 (7) ◽

pp. 910-913 ◽

Cited By ~ 5

Author(s):

Randall P. Niedz ◽

Michael G. Bausher ◽

C. Jack Hearn

Keyword(s):

Gel Electrophoresis ◽

Wheat Germ ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Carbohydrate Binding ◽

Pvdf Membrane ◽

Polyvinylidene Difluoride ◽

Sds Page ◽

Background Staining ◽

Avidin Biotin Complex

leaf and seed glycoproteins were detected after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotting onto polyvinylidene difluoride (PVDF) membrane when probed with biotinylated lectin at 1 μg·ml-1. Four lectins representing three carbohydrate-binding groups were used as probes. A preformed avidin-biotin-complex (ABC) was used to detect the glycoprotein-bound lectins and resulted in dark bands and little background staining. Concanavalin A (ConA) and wheat germ agglutinin (WGA) resulted in the darkest-staining bands. The four Citrus spp. and one related species studied had unique seed glycoprotein profiles when probed with ConA and WGA. This procedure might be useful in clarifying citrus taxonomy, providing genetic markers, and in physiological studies involving glycoproteins.

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