SDS-PAGE and Western Blot Analysis

Cell lysates or gradient centrifugation fractions were analysed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot as previously described (Troese et al., 2011 (link)). Commercially available primary antibodies targeted β-actin (Santa Cruz Biotechnology, Santa Cruz, CA), GAPDH (Sigma-Aldrich), GFP (Invitrogen), HaloTag (Promega), TGN46 (Sigma-Aldrich) and Rab10 (Sigma-Aldrich). Rabbit antisera against A. phagocytophilum proteins P44 (IJdo et al., 1999 (link)) and APH0032 (Huang et al., 2010b (link)) were described previously. Rabbit antibody targeting a surface-exposed epitope of Asp55 (Ge and Rikihisa, 2007 (link)) was a kind gift from Yasuko Rikihisa (Ohio State University, Columbus, OH).

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Truchan H.K., VieBrock L., Cockburn C.L., Ojogun N., Griffin B.P., Wijesinghe D.S., Chalfant C.E, & Carlyon J.A. (2015). Anaplasma phagocytophilum Rab10-dependent parasitism of the trans-Golgi network is critical for completion of the infection cycle. Cellular microbiology, 18(2), 260-281.

Publication 2015

β-actin GAPDH TGN46 Rab10

Actin Antibodies Antibody Antisera Cell Centrifugation Epitope Gapdh Halotag Promega Proteins a Rabbit Sds page Western blot

Corresponding Organization : Virginia Commonwealth University

Other organizations : Institute for Molecular Medicine, Hunter Holmes McGuire VA Medical Center

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Variable analysis

independent variables

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE)
Western blot

dependent variables

Protein expression levels of β-actin, GAPDH, GFP, HaloTag, TGN46, Rab10, P44, APH0032, and Asp55

control variables

Cell lysates or gradient centrifugation fractions
Commercially available primary antibodies
Rabbit antisera against A. phagocytophilum proteins
Rabbit antibody targeting a surface-exposed epitope of Asp55

positive controls

Rabbit antisera against A. phagocytophilum proteins P44 and APH0032
Rabbit antibody targeting a surface-exposed epitope of Asp55

negative controls

Not explicitly mentioned

Annotations

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