PHA-L Lectin Binding Assay

1 × 10⁶ cells from each cell line were harvested, washed twice with PBS, re-suspended in staining buffer (PBS + 0.2% FBS) and blocked (FcR Blocking Reagent, Miltenyi Biotec). Cells were then incubated with PHA-L lectin (GlycoMatrix) at a concentration of 10 µg/mL for 30 min at 4 °C. Following incubation, cells were washed twice with PBS, and re-suspended in staining buffer. 30,000 events were recorded for each tube using a BD LSRII Cell Analyzer. All data was further processed using FlowJo software.

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Butler W., McDowell C., Yang Q., He Y., Zhao Y., Hauck J.S., Zhou Y., Zhang H., Armstrong A.J., George D.J., Drake R, & Huang J. (2023). Rewiring of the N-Glycome with prostate cancer progression and therapy resistance. NPJ Precision Oncology, 7, 22.

Publication 2023

FcR Blocking Reagent BD LSRII Cell Analyzer

Buffer Cell Cells line Lectin Pha l

Corresponding Organization :

Other organizations : Duke University, Medical University of South Carolina, First Hospital of China Medical University, China Medical University, Dana-Farber Cancer Institute

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Variable analysis

independent variables

PHA-L lectin concentration (10 µg/mL)

dependent variables

Flow cytometry analysis of cells (30,000 events recorded for each tube)

control variables

Cell count (1 × 10^6 cells from each cell line)
Washing cells with PBS
Resuspending cells in staining buffer (PBS + 0.2% FBS)
Blocking with FcR Blocking Reagent
Incubation time (30 min)
Incubation temperature (4 °C)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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