Approximately 3 ml of venous blood (AA and SS) was collected in EDTA, centrifuged, and erythrocytes isolated. Approximately 2 million erythrocytes were incubated with 10μg of rhSiglec-9 protein (R&D Systems) for 30 minutes on ice. Erythrocytes were then stained with either anti-CD235a (Biolegend) or anti-IgG (Biolegend) on ice for 30 minutes and analyzed by flow cytometry. One set of AA erythrocytes were pretreated with 20mU of Arthrobacter ureafaciens sialidase (AUS) (Sigma-Aldrich) for 15 minutes at RT followed by protein incubation and staining [16 (link)].
Anti cd235a
Manufactured by BioLegend
Anti-CD235a is a laboratory reagent used to detect and analyze cells expressing the CD235a antigen, also known as glycophorin A. CD235a is a sialoglycoprotein found on the surface of red blood cells and their precursors. This reagent can be used in flow cytometry and other immunoassays to identify and quantify red blood cells and related cell populations.
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2 protocols using anti cd235a
1
Siglec-9 Binding Assay on Erythrocytes
2
Monoclonal Antibodies for Cell Surface Marker Analysis
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Rat monoclonal anti‐SSEA3 (Clone MC‐631) along with the following mouse monoclonal antibodies were used: anti‐SSEA3, anti‐SSEA4 (Clone MC‐813–70), anti‐CD34 (Clone V MA27), anti‐CD41 (Integrin GPIIb, Clone IV P38), anti‐CD235ab (Glycophorin A/B, Clone VII 70299), anti‐CD235a (Glycophorin A, Clone VII 70312), anti‐CD71 (transferrin receptor, Clone A015) (Biolegend, San Diego, CA); anti‐CD238 (Kell, Clone BRIC 203), anti‐CD234 (Duffy, Clone 2C3) (BD Biosciences, San Jose, CA); anti‐RhAG (Clone LA1818) (kindly provided by Prof Ellen van der Schoot, Sanquin, Amsterdam, Netherlands); anti‐RhD/CE (Clone BRIC69, Fisher Scientific); anti‐U (Glycophorin B, Human plasma, Versiti‐Wisconsin); anti‐CD233 (BAND3, Clone BRIC6, American Research Products, Palos Verdes Estates, CA); anti‐CD173 (H antigen, Clone BRIC 231, MyBioSource, San Diego, CA). Secondary antibodies used were as follows: monoclonal APC‐conjugated Rat anti‐mouse IgG1 (Clone RMG1‐1, Biolegend) and polyclonal APC‐conjugated goat anti‐human IgG (Fab) (RRID: AB_2337691, Jackson Laboratories, Bar Harbor, ME).
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