Flow Cytometric Analysis of Procyclin Expression

To monitor the proportion of the population expressing EP and/or GPEET procyclins, flow cytometry was performed as described previously [19 (link),20 (link)]. The primary antibodies were anti-EP (mouse monoclonal TBRP1/247, Cedarlane Laboratories, Burlington, Canada) and anti-GPEET (rabbit polyclonal K1) [79 (link)] and the secondary antibodies were Alexa Fluor 488-conjugated goat anti-rabbit (Invitrogen, Eugene, Oregon, USA) and Cy3-conjugated goat anti-mouse (Jackson ImmunoResearch). Antibody dilutions are mentioned in S3 Table. Fluorescence of 20,000 cells was measured using a Novocyte flow cytometer system (ACEA Biosciences, Inc., San Diego, USA).

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Bevkal S., Naguleswaran A., Rehmann R., Kaiser M., Heller M, & Roditi I. (2021). An Alba-domain protein required for proteome remodelling during trypanosome differentiation and host transition. PLoS Pathogens, 17(1), e1009239.

Publication 2021

Alexa Fluor 488-conjugated goat anti-rabbit Cy3-conjugated goat anti-mouse Novocyte flow cytometer system

Alexa fluor 488 Antibodies Antibody Cells Dilutions Flow cytometry Fluorescence Goat Gpeet procyclins Mouse Rabbit

Corresponding Organization : University of Basel

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Variable analysis

independent variables

Proportion of the population expressing EP procyclins
Proportion of the population expressing GPEET procyclins

dependent variables

Fluorescence of 20,000 cells

control variables

Flow cytometry protocol as described in previous studies [19,20]
Antibody dilutions as mentioned in S3 Table

controls

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

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