Biotinylated rca 1

Manufactured by Vector Laboratories

Biotinylated RCA-I is a plant lectin derived from the seeds of the castor oil plant (Ricinus communis). It binds specifically to galactose and N-acetylgalactosamine residues on cell surfaces. Biotinylation allows for the detection and visualization of glycoconjugates in various applications.

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Lab products found in correlation

Protease inhibitor, by Roche (1 mentions) Neuraminidase, by Merck Group (1 mentions) Ecl reagent, by Cytiva (1 mentions)

Close spelling variants of this product

Biotinylated RCA RCA-I

2 protocols using biotinylated rca 1

C3H10T1/2 Cell Glycosylation Analysis

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The C3H10T1/2 cells were collected at the indicated time, washed with PBS, scraped off, and collected by centrifugation, and then lysed in RIPA buffer (50 mM Tris-HCl [pH 8.0], 150 mM NaCl,1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) in the presence of protease inhibitors (Roche) at 4 °C. Cell lysates proteins were separated by SDS–PAGE and transferred onto a PVDF membrane. The PVDF membrane was then incubated in 25 mM H₂SO₄ at 4 °C for 1 h to eliminate terminal sialic acid moieties. After being blocked with 5% BSA, the membrane was immunoblotted with biotinylated RCA-I (Vector Laboratories, Burlingame, CA; 1:500 dilutions) for 2 h at room temperature and visualized with horseradish peroxidase-coupled avidin reagent.

Li S.F., Zhu C.S., Wang Y.M., Xie X.X., Xiao L.L., Zhang Z.C., Tang Q.Q, & Li X. (2018). Downregulation of β1,4-galactosyltransferase 5 improves insulin resistance by promoting adipocyte commitment and reducing inflammation. Cell Death & Disease, 9(2), 196.

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Sialic Acid Detection in Mouse Fibroblasts

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150 mm tissue culture plates of mouse fibroblasts were washed three times and lysed with 2 ml of 50 mM Na acetate, 2 mM CaCl₂, and 1% Triton. Particulate material was pelleted, the supernatant transferred to a clean tube for protein determination by Bradford reagent, and heat- inactivated for 10 min at 56 °C. Some samples were treated with 200 mU of neuraminidase (#N5631, Sigma) for 1 hr at 37 °C. Ten μg protein was resolved by 10% SDS-PAGE, transferred to nitrocellulose, and blocked overnight in TBST + 1% gelatin at 4 °C. Blots were incubated with biotinylated-RCA I [Vector] (2 μg/ml blocking buffer) for 1 h, followed by HRP-Streptavidin (1:50,000 blocking buffer) for 45 min. Blots were washed three times and developed with ECL Reagent (Amersham). As indicated, some blots were pre-incubated with 300 mM galactose for 10 min prior to incubation with RCA I.

Elder B.H, & Shur B.D. (2016). Mouse fibroblasts null for the long isoform of β1,4-galactosyltransferase-I show defective cell-matrix interactions. Biochemical and biophysical research communications, 478(3), 1248-1253.

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