Immunofluorescence Staining of Lipid Droplet Proteins

Rabbit polyclonal antibodies used: anti-GPAT4 (Wilfling et al., 2013 (link)), anti-CCT1 (Wilfling et al., 2013 (link)), anti-GBF1 (BD Biosciences, San Jose, CA), anti-KDEL-receptor (KDELR; gift from Dr JE Rothman; Yale University), anti-βCOP (gift from Dr JE Rothman; Yale University), anti-perilipin3 (TIP47; Novus Biologicals, Littleton, CO), anti-αCOP (Abcam, Cambridge, MA), anti-GRP78/BiP (ET-21) (Sigma–Aldrich, St. Louis, MO) and anti-garz (Wang et al., 2012 (link)) (gift from Dr A Paululat; University of Osnabrück). Mouse monoclonal antibodies used: anti-GM130 (BD Biosciences), anti-tubulin (Sigma–Aldrich), anti-β’COP (gift from Dr JE Rothman; Yale University), and anti-clathrin heavy chain (x22) (Thermo Scientific, Waltham, MA) antibody. The following secondary antibodies were used: Alexa Fluor 568 goat anti-rabbit (Invitrogen, Grand Island, NY), Alexa Fluor 488 goat anti-mouse (Invitrogen), ATTO 647N (STED) goat anti-rabbit (Active Motif, Carlsbad, CA), and goat anti-mouse STAR470SX (Abberior, Göttingen, Germany).

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Wilfling F., Thiam A.R., Olarte M.J., Wang J., Beck R., Gould T.J., Allgeyer E.S., Pincet F., Bewersdorf J., Farese RV J.r, & Walther T.C. (2014). Arf1/COPI machinery acts directly on lipid droplets and enables their connection to the ER for protein targeting. eLife, 3, e01607.

Publication 2014

anti-GBF1 anti-βCOP anti-GM130 anti-tubulin Alexa Fluor 568 goat anti-rabbit Alexa Fluor 488 goat anti-mouse

Alexa 568 Alexa fluor 488 Antibodies Antibody Biologicals Clathrin heavy chain Goat Kdel receptor Monoclonal antibodies Mouse Novus Rabbit Tubulin

Corresponding Organization :

Other organizations : Yale University, Sorbonne Université, Université Paris Cité, Heidelberg University, University of California, San Francisco, Gladstone Institutes

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Variable analysis

independent variables

Anti-GPAT4 antibody
Anti-CCT1 antibody
Anti-GBF1 antibody
Anti-KDEL-receptor (KDELR) antibody
Anti-βCOP antibody
Anti-perilipin3 (TIP47) antibody
Anti-αCOP antibody
Anti-GRP78/BiP (ET-21) antibody
Anti-garz antibody
Anti-GM130 antibody
Anti-tubulin antibody
Anti-β'COP antibody
Anti-clathrin heavy chain (x22) antibody
Alexa Fluor 568 goat anti-rabbit secondary antibody
Alexa Fluor 488 goat anti-mouse secondary antibody
ATTO 647N (STED) goat anti-rabbit secondary antibody
Goat anti-mouse STAR470SX secondary antibody

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

Annotations

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